5,6-dihydro-1,2,4-triazine compounds and their pharmaceutical use as glp-1 receptor agonists

Abstract

The application discloses a 5,6-dihydro-1,2,4-triazine compound and a pharmaceutical application of the 5,6-dihydro-1,2,4-triazine compound as a GLP-1 receptor agonist. The structure of the 5,6-dihydro-1,2,4-triazine compound is shown in a formula I, wherein definitions of the substituents are described in the description and the claims. The compound of the application can be used for treating diabetes and metabolic syndrome and the like as a GLP-1 receptor agonist.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, specifically to a 5,6-dihydro-1,2,4-triazine compound, its preparation method, and its pharmaceutical applications. The invention further relates to the use of said compound, its salt or solvate, and pharmaceutical compositions containing the compound as GLP-1 receptor agonists in the treatment of diabetes and metabolic syndrome. Background Technology

[0002] Diabetes is the third leading cause of death among chronic diseases threatening human health, after cancer and cardiovascular diseases. According to the eighth edition of the International Diabetes Federation's (IDF) Global Diabetes Atlas, there are currently over 460 million people with diabetes worldwide, and this number is projected to reach nearly 700 million by 2045, a situation that is far from optimistic. As a multifactorial metabolic disease, diabetes is characterized by chronic hyperglycemia, accompanied by disorders of glucose, lipid, and protein metabolism caused by defects in insulin secretion or action. Two main forms of diabetes have been identified: type 1 diabetes (insulin-dependent diabetes mellitus, IDDM) and type 2 diabetes (non-insulin-dependent diabetes mellitus, NIDDM). Type 2 diabetes accounts for more than 90% of all diabetes cases and is characterized by decreased pancreatic β-cell function and insulin resistance in peripheral tissues such as the liver, skeletal muscle, and adipose tissue, leading to disorders of glucose and lipid metabolism. Other related diseases include hepatic insulin resistance, glucose intolerance, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, obesity, dyslipidemia, hypertension, hyperinsulinemia, and non-alcoholic fatty liver disease (NAFLD). In the past decade, three novel hypoglycemic agents have been clinically applied in type 2 diabetes mellitus (T2DM): glucagon-like peptide-1 (GLP-1) receptor agonists, dipeptidyl peptidase-4 (DPP-4) inhibitors, and sodium-glucose cotransporter 2 (SGLT2) inhibitors. Among these, GLP-1R agonists offer advantages such as potent glucose-dependent hypoglycemic effects, no increase in the risk of hypoglycemia, weight reduction, mild blood pressure reduction, and cardiovascular benefits, resulting in a significant increase in their market share in recent years. Increasing clinical trials have also demonstrated the significant effects of GLP-1R agonists in non-alcoholic fatty liver disease, improving cardiovascular outcomes, kidney protection, diabetic nephropathy, and Alzheimer's disease.

[0003] GLP-1 receptors are members of the G protein-coupled receptor (GPCR) class B family of peptide hormones, distributed throughout the body, including pancreatic islet cells, lungs, kidneys, brain, hypothalamus, cardiovascular system, gastrointestinal tract, skin, and vagus nerve. GLP-1 receptors may also be found in the liver, adipose tissue, and skeletal muscle. Its natural agonist ligand is GLP-1 (glucagon-like peptide-1), a peptide hormone encoded by the human glucagon gene and secreted by intestinal L cells, belonging to the incretin family. Physiologically, after ingestion of nutrients through food, GLP-1 is released from intestinal L cells into circulation at low picomol levels (5 to 15 pmol / L). These levels of GLP-1 help activate pancreatic β-cell GLP-1R to stimulate insulin secretion in a glucose-dependent manner, while simultaneously inhibiting glucagon secretion, thus lowering and maintaining a constant postprandial blood glucose level. GLP-1 also has neuroregulatory functions, reducing appetite, inhibiting gastric emptying, and promoting the growth of pancreatic β-cells. 50%-70% of orally ingested glucose in the human body is metabolized through the GLP-1 / insulin pathway, making GLP-1 an important hypoglycemic peptide. In terms of treatment, GLP-1 secretion levels are significantly reduced in patients with type 2 diabetes, which is one of the important reasons for uncontrolled blood glucose. Therefore, the development of GLP-1 analogs has been an important research direction for the treatment of type 2 diabetes. Endogenous GLP-1 can be rapidly degraded and inactivated by dipeptidyl peptidase-4 (DPP-4) in the body, with a very short half-life of approximately 1-2 minutes. Therefore, the body's own GLP-1 is not suitable for clinical treatment of diabetes. Currently, marketed GLP-1R agonist drugs and drugs under investigation mainly focus on peptide analogs.

[0004] Exenatide, a GLP-1 mimic, was the first GLP-1R agonist approved for the treatment of type 2 diabetes. Exenatide is equivalent to natural GLP-1 in activating GLP-1R, demonstrating effective glycemic control and weight loss at plasma concentrations ranging from 40 to 70 pmol / L. Other potent GLP-1R agonists, including liraglutide, duraglutide, and semaglutide, have been approved and offer cardiovascular health benefits. While these drugs provide better treatment outcomes, they still account for a limited percentage of diabetes prescriptions, largely because they are based on large peptide molecules and require invasive subcutaneous injection, leading to poorer patient adherence compared to oral administration. A breakthrough in GLP-1 therapy is the development of semaglutide tablets formulated with the absorption enhancer sodium N-(8-[2-hydroxybenzoyl]amino)octanoate for oral administration, with an oral bioavailability of less than 1%. Furthermore, because drug absorption is significantly affected by food and gastric juices, the oral administration regimen of semaglutide is limited. Specifically, tablets must be taken with a certain amount of water after an overnight fast and at least 30 minutes before breakfast or other medications. Therefore, small-molecule oral GLP-1R agonists, as an alternative, offer a more standardized drug formulation and a simpler administration method. Simultaneously, small molecules more easily cross the blood-brain barrier, acting on GLP-1R in the arcuate nucleus of the hypothalamus to reduce appetite, potentially leading to better weight loss while lowering blood sugar.

[0005] Currently, small molecule GLP-1R agonists under clinical research include TTP-273 (Hangzhou Sino-American East China Pharmaceutical, Vtv Therapeutics), LY-3502970 (Chung-Wai Pharmaceutical / Eli Lilly), and Danuglipron (PF-06882961, Pfizer). Among them, the Phase I clinical trial results of PF-06882961 showed significant effects in lowering blood sugar and reducing weight in patients with type 2 diabetes. In patients with type 2 diabetes, combined with metformin and twice-daily administration of PF-06882961 for 28 days, a dose of 120 mg reduced HBA1c by 1.2% and reduced weight by 7.9 kg (ClinicalTrials.govIdentifier:NCT03538743). A Phase II clinical trial is underway, attracting widespread attention from the pharmaceutical industry and research institutions. Further extensive and in-depth research on GLP-1R agonists is needed in this field. Summary of the Invention

[0006] The purpose of this invention is to provide a small molecule compound suitable for oral administration with superior physicochemical properties and better GLP-1R agonist activity for the prevention or treatment of diseases or symptoms related to downstream GLP-1R signaling pathways.

[0007] A first aspect of the present invention provides a compound of general formula (I) or a stereoisomer thereof, a deuterated thereof, or a pharmaceutically acceptable salt thereof:

[0008]

[0009] In the formula, ring A is a phenyl or a 5-6 membered heteroaryl group;

[0010] R 1 Selected from the group consisting of: 3-6-membered heterocyclic substituted C1-C6 alkyl, C3-C6 cycloalkyl substituted C1-C6 alkyl, 5-8-membered heteroaryl substituted C1-C6 alkyl, C1-C6 alkoxy substituted C1-C6 alkyl, 3-6-membered heterocyclic, C3-C6 cycloalkyl, or 5-8-membered heteroaryl; wherein the 3-6-membered heterocyclic, C3-C6 cycloalkyl, 5-8-membered heteroaryl, or C1-C6 alkoxy group is optionally substituted by one or more substituents selected from the group consisting of: H, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 Alkoxy, C1-C6 alkylamino, cyano, cyano-substituted C1-C6 alkyl, hydroxy, hydroxy-substituted C1-C6 alkyl, halo-C1-C6 alkyl, halo-C3-C6 cycloalkyl, halo-C1-C6 alkoxy, halo-C1-C6 alkylamino, acyl, amide, aminoacyl, sulfonyl, amino.

[0011] R 2 and each R 3 Each is independently H, halogen, or C1-C6 alkyl; or 2 R 3 Linkage forms C3-C8 cycloalkyl or 4-8 membered heterocyclic groups; each R 4 Independently H, halogen, or C1-C6 alkyl; R 5 It is H or C1-C6 alkyl;

[0012] Q is

[0013] Among them, Z 1 Z 2 Z 3 and Z 4 Each independently for CR 11 Or N; Y is O or S;

[0014] Ring B is a C6-C10 aryl, 5-8 membered heteroaryl, or The ring C is a 5-8 membered heteroaryl, a C5-C8 partially unsaturated cycloalkyl, or a 5-8 membered partially unsaturated heterocyclic group; W 1 and W 2 Each independently selected from CR 12 Or N;

[0015] Each R6 Independently selected from the group consisting of: H, halogen, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic, halo-C1-C6 alkyl, halo-C1-C6 alkoxy, C3-C8 cycloalkyl-substituted C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, C1-C6 alkylamino-substituted C1-C6 alkyl, C6-C10 aryl, 5-8 membered heteroaryl, nitro, cyano, cyano-substituted C1-C6 alkyl, hydroxyl, -SR, -N(R)2, -C(O)OR, -C(O)N(R)2, -C(O)R, -S(O)R, -S(O)2R, -S(O)2N(R)2 or -N(R)C(O)R; each R is independently selected from the following group: H, C1-C6 alkyl, C3-C8 cycloalkyl, halo-C1-C6 alkyl, hydroxy-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl or C1-C6 alkylamino-substituted C1-C6 alkyl;

[0016] R 7 It is H, C1-C6 alkyl, or halo-C1-C6 alkyl;

[0017] Each R 8 It can be independently H, halogen, cyano, C1-C6 alkyl, or halo-C1-C6 alkyl;

[0018] R 9 It can be H, halogen, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, halo-C1-C6 alkyl, halo-C1-C6 alkoxy, cyano or cyano-substituted C1-C6 alkyl;

[0019] Each R 10 Independently H, halogen, C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkyl or halo-C1-C6 alkoxy, or 2 R 10 Linkage forms C3-C8 cycloalkyl, 4-8 membered heterocyclic groups, or phenyl groups;

[0020] Each R 11 Independently, each R is H, halogen, cyano, C1-C6 alkyl, or halo-C1-C6 alkyl. 12 Independently, it is H, halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkyl, or halo-C1-C6 alkoxy;

[0021] a is 0, 1, 2, 3, or 4; m is 0, 1, 2, or 3; n is 0, 1, 2, 3, 4, or 5;

[0022] t is 0, 1, 2 or 3; p is 0, 1, 2, 3 or 4.

[0023] In another preferred embodiment, Z 1 Z 2 Z 3 and Z 4 Each independently for CR 11 Y is O; ring B is C6 aryl.

[0024] In another preferred embodiment, ring A is phenyl, thienyl, or pyridyl.

[0025] In another preferred embodiment, the compound has the structure shown in formula (II) or formula (III):

[0026]

[0027] Where: R 1 R 2 R 3 R 4 R 5 Q, a, and m are as defined above.

[0028] In another preferred embodiment, Z 1 Z 2 and Z 3 Each independently selected from CR 11 Or N, Z 4 For N; R 11 As mentioned above.

[0029] In another preferred embodiment, Z 1 Z 2 and Z 3 Each can be independently CH, CF, CCl, or N.

[0030] In another preferred embodiment, Q is selected from Q3-Q11.

[0031] In another preferred embodiment, Selected from the following group: Among them, R 6 R 9 R 10 n and p are as defined above.

[0032] In another preferred embodiment, the ring C is selected as a 5-6 membered heteroaryl or a 5-6 membered heterocyclic group.

[0033] In another preferred embodiment, for

[0034] In another preferred embodiment, for

[0035] In another preferred embodiment, the compound has the structure shown in formula (IV), formula (V), formula (VI) or formula (VII).

[0036] In another preferred embodiment, R 1 Selected from the group consisting of: 4-6-membered heterocyclic substituted C1-C4 alkyl, C3-C6 cycloalkyl substituted C1-C4 alkyl, 5-6-membered heteroaryl substituted C1-C4 alkyl, C1-C4 alkoxy substituted C1-C4 alkyl, 4-6-membered heterocyclic, C3-C6 cycloalkyl, or 5-6-membered heteroaryl; wherein the 4-6-membered heterocyclic, C3-C6 cycloalkyl, 5-6-membered heteroaryl, or C1-C4 alkoxy group is optionally substituted by one or more substituents selected from the group consisting of: H, halogen, C1-C4 alkyl, C3-C6 cycloalkyl, C1-C4 alkoxy, C1-C4 alkylamino, cyano, cyano-substituted C1-C4 alkyl, hydroxy, hydroxy-substituted C1-C4 alkyl, halo-C1-C4 alkyl, halo-C3-C6 cycloalkyl, halo-C1-C4 alkoxy, halo-C1-C4 alkylamino, acyl, amide, aminoacyl, sulfonyl, amino.

[0037] In another preferred embodiment, R 1 Selected from 4-6-membered heterocyclic substituted C1-C4 alkyl, 5-6-membered heteroaryl substituted C1-C4 alkyl, and C3-C6 cycloalkyl substituted C1-C4 alkyl; wherein the above 4-6-membered heterocyclic, C3-C6 cycloalkyl, and 5-6-membered heteroaryl groups are optionally substituted by one or more substituents selected from the group consisting of: halogen, C1-C4 alkyl, C3-C6 cycloalkyl, C1-C4 alkoxy, C1-C4 alkylamino, cyano, cyano-substituted C1-C4 alkyl, hydroxyl, and hydroxyl-substituted C1-C4 alkyl.

[0038] In another preferred embodiment, R 1 The group is selected from C1 alkyl groups substituted with 4-membered heterocyclic groups, C1 alkyl groups substituted with 5-6-membered heteroaryl groups, and C1-C4 alkyl groups substituted with C3 cycloalkyl groups; the above-mentioned 4-membered heterocyclic groups, C3 cycloalkyl groups, and 5-6-membered heteroaryl groups may be substituted by one or more substituents selected from the group consisting of: halogen, C1-C4 alkyl, C3-C6 cycloalkyl, cyano, and cyano-substituted C1-C4 alkyl groups.

[0039] In another preferred embodiment, R 1 Selected from

[0040] In another preferred embodiment, R 2 and each R 3 Each is independently selected from H or C1-C3 alkyl groups; preferably R.2 and R 3 Each is independently selected from H, methyl, or ethyl.

[0041] In another preferred embodiment, R 4 Each is independently selected from H, halogens, or C1-C3 alkyl groups; preferably R. 4 It can be H, Cl, or F.

[0042] In another preferred embodiment, R 5 Each is independently selected from H or C1-C3 alkyl groups; preferably R. 5 It is H, methyl, or ethyl. In another preferred embodiment, R 6 Each is independently selected from H, halogen, C1-C3 alkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy, and cyano; preferably R. 6 It can be H, F, Cl, cyano, methoxy, trifluoromethyl, difluoromethyl, or difluoromethoxy.

[0043] In another preferred embodiment, R 7 Selected from H or C1-C3 alkyl groups; preferably R 7 It is H or methyl.

[0044] In another preferred embodiment, each R 8 Independently H, halogen, or C1-C3 alkyl; preferably R 8 For H.

[0045] In another preferred embodiment, R 9 Selected from H, halogen, C1-C3 alkyl, halogen-substituted C1-C3 alkyl, or cyano; preferably R. 9 It can be F, Cl, cyano, trifluoromethyl or difluoromethyl.

[0046] In another preferred embodiment, R 10 Each is independently selected from H, halogen, C1-C3 alkyl, or halogen-substituted C1-C3 alkyl; preferably R. 10 It consists of H, F, and methyl groups.

[0047] In another preferred embodiment, R 11 Each is independently selected from H, halogens, or C1-C3 alkyl groups; preferably R. 11 It can be H or F.

[0048] In another preferred embodiment, R 12 Each is independently selected from H, halogens, or C1-C3 alkyl groups; preferably R. 12 For H.

[0049] In another preferred embodiment, R 1 R 2 R 3 R4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 The Q ring, B ring, X, and Y are the corresponding functional groups in the compounds prepared in the examples.

[0050] In another preferred embodiment, the compounds are selected from: compounds 1-59, compounds 61-63.

[0051] The compounds of this invention have an asymmetry center, a chiral axis, and a chiral plane, and can exist as racemates, R-isomers, or S-isomers. Those skilled in the art can obtain the R-isomers and / or S-isomers from the racemates using conventional techniques.

[0052] In another preferred embodiment, the pharmaceutically acceptable salt of the present invention is a salt formed by an anion and a positively charged group on a compound of general formula (I). Suitable anions are chloride, bromide, iodide, sulfate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, acetate, malate, toluenesulfonate, tartrate, fumarate, glutamate, glucuronide, lactate, glutarate, or maleate. Similarly, salts can be formed by cations and negatively charged groups on compounds of general formula (I). Suitable cations include sodium, potassium, magnesium, calcium, and ammonium ions.

[0053] In another preferred embodiment, the pharmaceutically acceptable salt of the present invention refers to a salt formed by a compound of general formula (I) with an acid from the following group: hydrofluoric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, nitric acid, methanesulfonic acid, aminosulfonic acid, salicylic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, citric acid, acetic acid, lactic acid, tartaric acid, succinic acid, oxalic acid, pyruvic acid, malic acid, glutamic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, and ethanesulfonic acid. Naphthalene disulfonic acid, malonic acid, fumaric acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pyric acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, and hydroxyethanesulfonic acid, etc.; or sodium, potassium, calcium, aluminum, or ammonium salts formed by compounds of general formula (I) with inorganic bases; or methylamine, ethylamine, or ethanolamine salts formed by compounds of general formula (I) with organic bases.

[0054] In another preferred embodiment, the pharmaceutically acceptable salt is a sodium salt.

[0055] In another preferred embodiment, the pharmaceutically acceptable salt is an ammonium salt.

[0056] In another preferred embodiment, the pharmaceutically acceptable salt is 1,3-dihydroxy-2-(hydroxymethyl)propyl-2-amine salt.

[0057] A second aspect of the present invention provides a pharmaceutical composition comprising:

[0058] The compound or its stereoisomer, its deuterated form or its pharmaceutically acceptable salt described in the first aspect; and a pharmaceutically acceptable carrier, diluent or excipient.

[0059] The compounds and pharmaceutical compositions provided by this invention can be in various forms, such as tablets, capsules, powders, syrups, solutions, suspensions, and aerosols, and can be contained in suitable solid or liquid carriers or diluents. The pharmaceutical compositions of this invention can also be stored in suitable sterile injection or infusion apparatus. The pharmaceutical compositions may also contain odorants, flavorings, etc.

[0060] This invention provides novel compounds that can be used alone or mixed with pharmaceutically acceptable excipients (e.g., excipients, diluents, etc.) to formulate oral tablets, capsules, granules, or syrups. The pharmaceutical composition can be prepared using conventional pharmaceutical methods.

[0061] In another preferred embodiment, the pharmaceutical composition further comprises at least one other therapeutic agent. Preferably, the other therapeutic agent is a diabetes treatment agent, a cardiovascular treatment agent, or an obesity treatment agent.

[0062] In this invention, the pharmaceutical composition contains a safe and effective amount (e.g., 0.1-99.9 parts by weight, preferably 1-90 parts by weight) of a compound of general formula (I) or a pharmaceutically acceptable salt thereof; and the balance being a pharmaceutically acceptable excipient, wherein the total weight of the composition is 100 parts by weight. Alternatively, the pharmaceutical composition of this invention contains 0.1-99.9% by weight, preferably 1-90% by weight, of a compound of general formula (I) or a pharmaceutically acceptable salt thereof; and the balance being a pharmaceutically acceptable excipient, wherein the total weight of the composition is 100% by weight.

[0063] The preferred ratio of the compound represented by general formula (I) to a pharmaceutically acceptable carrier, excipient or sustained-release agent is such that the compound represented by general formula (I) constitutes more than 60% of the total weight as the active ingredient, and the remainder constitutes 0-40% of the total weight, preferably 1-20%, and most preferably 1-10%.

[0064] A third aspect of the invention provides the use of the compound described in the first aspect or its stereoisomer, its deuterated form or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the second aspect, for the preparation of a GLP-1 receptor agonist; or for the preparation of a medicament for the prevention and / or treatment of diseases or symptoms related to the GLP-1R downstream signaling pathway.

[0065] In another preferred embodiment, the diseases or symptoms associated with the downstream signaling pathway of GLP-1R are selected from: diabetes, metabolic syndrome, diabetic complications, obesity, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), Parkinson's disease, dementia, hyperglycemia, impaired glucose tolerance, atherosclerosis, hypertension, hyperlipidemia, coronary heart disease, cerebral infarction or stroke; preferably type II diabetes, obesity, diabetic complications, non-alcoholic steatohepatitis or cardiovascular disease.

[0066] The compounds of general formula (I) of this invention are characterized by a novel 5,6-dihydro-1,2,4-triazine ring as the connecting heterocycle. Some compounds exhibit GLP-1 receptor agonist activity comparable to, or even higher than, that of the control compound PF-06882961. Unexpectedly, the introduction of the 5,6-dihydro-1,2,4-triazine ring significantly weakens the blocking effect of the compounds on hERG channels, and is expected to have a lower risk of cardiotoxicity. Furthermore, when administered orally, the compounds of this invention can significantly reduce blood glucose concentration and food intake, showing potential for the treatment of diabetes and obesity.

[0067] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0068] Figure 1 The blood glucose curves of compound 10 and the positive control PF-06882961 in a mouse glucose tolerance test are shown.

[0069] Figure 2 The area under the curve (AUC) of compound 10 and the positive control PF-06882961 in a mouse glucose tolerance test is shown. (0-120min) .

[0070] Figure 3 The food intake of compound 10 and the positive control PF-06882961 in a mouse feeding experiment is shown. Detailed Implementation

[0071] The inventors of this application, through extensive and in-depth research, developed a 5,6-dihydro-1,2,4-triazine compound as a GLP-1 receptor agonist, which can be used to treat diseases such as diabetes and metabolic syndrome. Based on this, the present invention was completed.

[0072] the term

[0073] In this invention, unless otherwise specified, the terms used have the general meanings known to those skilled in the art.

[0074] In this invention, the halogen is F, Cl, Br or I.

[0075] In this invention, the terms "C1-C6" refer to having 1, 2, 3, 4, 5, or 6 carbon atoms, "C3-C8" refers to having 3, 4, 5, 6, 7, or 8 carbon atoms, and "C1-C3" refers to having 1, 2, or 3 carbon atoms, and so on. "3-6-membered" refers to having 3, 4, 5, or 6 ring atoms, "5-8-membered" refers to having 5, 6, 7, or 8 ring atoms, and so on.

[0076] In this invention, the term "alkyl" refers to a saturated linear or branched hydrocarbon moiety. For example, the term "C1-C6 alkyl" refers to a straight-chain or branched alkyl group having 1 to 6 carbon atoms, and includes, without limitation, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, and hexyl; preferably ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0077] In this invention, the term "alkoxy" refers to an -O-(C1-6 alkyl) group. For example, the term "C1-C6 alkoxy" refers to a straight-chain or branched alkoxy group having 1 to 6 carbon atoms, and includes, without limitation, methoxy, ethoxy, n-propoxy, isopropoxy, and butoxy groups.

[0078] In this invention, the term "alkenyl" refers to a straight-chain or branched hydrocarbon moiety containing at least one double bond. For example, the term "C2-C6 alkenyl" refers to a straight-chain or branched alkenyl moiety containing one double bond and having 2 to 6 carbon atoms, and includes, without limitation, vinyl, propenyl, butenyl, isobutenyl, pentenyl, and hexenyl.

[0079] In this invention, the term "alkynyl" refers to a straight-chain or branched alkynyl group containing a triple bond, and includes, without limitation, ethynyl, propynyl, butynyl, isobutynyl, pentylyl, and hexynyl.

[0080] In this invention, the term "cycloalkyl" or "carbocyclic" refers to a saturated cyclic hydrocarbon moiety, such as the term "C3-C". 10"Cycloalkyl" refers to a cyclic alkyl group having 3 to 10 carbon atoms on a ring, and includes, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclodecyl. The terms "C3-C8 cycloalkyl", "C3-C7 cycloalkyl", and "C3-C6 cycloalkyl" have similar meanings.

[0081] In this invention, the term "aryl" refers to a hydrocarbon moiety comprising one or more aromatic rings. For example, the term "C6-C..." 12 "Aryl" refers to an aromatic cyclic group with 6 to 12 carbon atoms that does not contain heteroatoms on the ring, such as phenyl and naphthyl. The term "C6-C12 aryl" has a similar meaning. Examples of aryl groups include, but are not limited to, phenyl (Ph), naphthyl, pyrene, anthracene, and phenanthryl.

[0082] In this invention, the term "heterocyclic group" refers to a saturated or unsaturated, non-aromatic cyclic group containing at least one (e.g., 1, 2, 3, or 4) cyclic heteroatoms (e.g., N, O, or S), such as oxacyclobutyl, pyrrolidinyl, tetrahydropyridyl, pyrrolinyl, dihydropyridyl, dihydrofuranyl, dihydrothiophenyl, or morpholinyl.

[0083] In this invention, the term "heteroaryl" refers to an aromatic cyclic group containing at least one (e.g., 1, 2, 3, or 4) cyclic heteroatoms (e.g., N, O, or S), such as furanyl, pyrroleyl, thiopheneyl, oxazolyl, imidazolyl, thiazolyl, pyridinyl, quinolinyl, isoquinolinyl, indolyl, pyrimidinyl, and pyranyl.

[0084] Unless otherwise stated, the alkyl, alkoxy, cycloalkyl, heteroaryl, heterocyclic, and aryl groups mentioned herein are substituted and unsubstituted groups. Possible substituents on the alkyl, alkoxy, cycloalkyl, heterocyclic, and aryl groups include, but are not limited to: hydroxyl, amino, nitro, nitrile, halogen, C1-C6 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C20 cycloalkyl, ... C3-C20 cycloalkenyl, C3-C20 heterocyclic alkyl, C1-C20 heterocyclic alkenyl, C1-C6 alkoxy, C6-C10 aryl, heteroaryl, heteroaryloxy, C1-C10 alkylamino, C1-C20 dialkylamino, C6-C10 arylamino, diC6-C10 arylamino, C1-C10 alkylaminosulfonyl, C6-C10 arylaminosulfonyl, C1-C10 alkylimino, C1-C10 alkylsulfonylimino, C6-C10 arylsulfonylimino, mercapto, C1-C10 alkylthio, C1-C10 alkylsulfonyl, C6-C10 arylsulfonyl, acylamino, aminoacyl, aminothioacyl, guanidinyl, ureyl, cyano, acyl, thioacyl, acyloxy, carboxyl, and carboxylic acid ester. On the other hand, cycloalkyl, heterocycloalkyl, heterocycloalkenyl, aryl, and heteroaryl groups can also fused together.

[0085] In this invention, the substitution can be monosubstituted or polysubstituted, and the polysubstituted can be disubstituted, trisubstituted, tetrasubstituted, or pentasubstituted. Disubstituted means having two substituents, and so on. When the substitution is polysubstituted, the substituents can be the same or different.

[0086] In this invention, "optionally" means that the defined group can be selected from a series of candidate groups, or it can be left unselected.

[0087] In this invention, appropriate protecting groups and methods of protecting and deprotecting different substituents using such appropriate protecting groups are well known to those skilled in the art; examples of which are found in T. Greene and P. Wuts, Protecting Groups in Organic Synthesis (4th edition), John Wiley & Sons (2007), which are incorporated herein by reference in their entirety.

[0088] The definition of "pharmaceutically acceptable salt" in this invention is as described in the invention summary.

[0089] Pharmaceutical Composition

[0090] The present invention also provides a pharmaceutical composition comprising an active ingredient within a safe and effective range, and a pharmaceutically acceptable carrier, excipient, or sustained-release agent.

[0091] The "active ingredient" mentioned in this invention refers to the compound of formula I described in this invention.

[0092] The "active ingredient" and pharmaceutical composition described in this invention are used to prepare drugs that activate GLP-1 receptors or to prevent or treat diseases and / or symptoms associated with downstream signaling pathways of GLP-1R.

[0093] The diseases or symptoms associated with the downstream signaling pathway of GLP-1R are: diabetes, metabolic syndrome, diabetic complications, obesity, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), Parkinson's disease, dementia, hyperglycemia, impaired glucose tolerance, atherosclerosis, hypertension, hyperlipidemia, coronary heart disease, cerebral infarction or stroke; preferably type II diabetes, obesity, diabetic complications, non-alcoholic steatohepatitis or cardiovascular disease.

[0094] "Diabetic complications" are complications arising from diabetes or hyperglycemia, and they can be acute or chronic. The term "acute complications" includes ketoacidosis and infectious diseases (such as skin infections, soft tissue infections, biliary tract infections, respiratory infections, and urinary tract infections), while "chronic complications" include, for example, microvascular diseases (such as nephropathy and retinopathy), neuropathy (such as sensory nerve disorders, motor nerve disorders, and autonomic nerve disorders), and gangrene. Major diabetic complications include diabetic retinopathy, diabetic nephropathy, and diabetic neuropathy.

[0095] Coronary heart disease includes asymptomatic myocardial ischemia (silent coronary heart disease), myocardial infarction, angina pectoris, ischemic heart failure (ischemic cardiomyopathy), and sudden death.

[0096] "Dementia" includes, for example, Alzheimer's disease, early-onset dementia (EOD), vascular dementia, and diabetic dementia.

[0097] "Safe and effective dose" refers to an amount of active ingredient sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of active ingredient per dose, more preferably 10-200 mg of active ingredient per dose. Preferably, "one dose" refers to one tablet.

[0098] "Pharmaceutically acceptable carriers" refer to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here refers to the ability of the components in the composition to interact with and incorporate the active ingredient of the invention without significantly reducing the efficacy of the active ingredient. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as... Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0099] The administration method of the active ingredient or pharmaceutical composition of the present invention is not particularly limited, and representative administration methods include (but are not limited to): oral administration and intravenous, intramuscular, or subcutaneous administration. From the standpoint of ease of preparation and administration, the preferred pharmaceutical compositions are solid compositions, especially tablets and solid-filled or liquid-filled capsules. Oral administration of the pharmaceutical composition is preferred.

[0100] Compounds of general formula (I) or pharmaceutical compositions comprising compounds of general formula (I) may be used clinically in mammals, including humans and animals, via routes of administration including oral, nasal inhalation, transdermal absorption, pulmonary administration, or gastrointestinal administration. Oral administration is preferred. A single-dose or divided dose is preferred. Regardless of the method of administration, the optimal dose for an individual should be determined based on the specific treatment. Generally, a low dose is started, and the dose is gradually increased until the most suitable dose is found.

[0101] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. Solid carriers include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose, and kaolin, while liquid carriers include sterile water, polyethylene glycol, nonionic surfactants, and edible oils (such as corn oil, peanut oil, and sesame oil), provided they are suitable for the characteristics of the active ingredient and the desired specific route of administration. Adjuvants commonly used in the preparation of pharmaceutical compositions may also be advantageously included, such as flavoring agents, colorings, preservatives, and antioxidants such as vitamin E, vitamin C, BHT, and BHA.

[0102] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active ingredient, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures thereof. Besides these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0103] In addition to the active ingredient, the suspension may contain suspending agents, such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0104] Injectable formulations include, but are not limited to, sterile, injectable, aqueous, oil-containing solutions, suspensions, emulsions, etc. These formulations can also be formulated with suitable parenteral diluents, dispersants, wetting agents, suspending agents, etc. Such injectable formulations can be sterilized by filtration through a bacteria-retaining filter. These formulations can also be formulated with bactericides dissolved or dispersed in an injectable medium or using other methods known in the art.

[0105] The compounds of this invention can be administered alone or in combination with other known drugs for treating or improving similar conditions.

[0106] The compounds of the present invention can be used in combination with one or more other drugs to treat, prevent, or improve diseases for which the compounds of the present invention or other drugs may be effective, wherein the combination of these drugs is safer or more effective than the use of any one drug alone. The other drugs may be administered simultaneously with, before, or after the compounds of the present invention via the usual route of administration and dosage. When the compounds of the present invention are used simultaneously with one or more other drugs, a unit dosage form of a pharmaceutical composition comprising the other drug and the compound of the present invention is preferred. However, drug combination may also include therapies in which the compounds described herein and one or more other drugs are administered in different overlapping regimens. When used in combination with one or more other active ingredients, the compounds of the present invention and the other drugs may be used at lower doses than when used alone.

[0107] Drugs or active ingredients that can be used in combination with the compounds described in this invention include, but are not limited to, the following drugs for treating diabetes: biguanides, thiazolidinediones, megglitazones, sulfonylureas, DPP4 inhibitors, SGLT1 and / or SGLT2 inhibitors, GPR40 agonists, α-glucosidase inhibitors, insulin, insulin analogs, GIPR agonists, GLP-1 / GIP dual receptor agonists, GLP-1 / GCGR dual receptor agonists, GLP-1 / GCGR dual receptor agonists, or GLP-1 / GIP / GCGR triple receptor agonists.

[0108] Drugs or active ingredients that can be used in combination with the compounds described in this invention include, but are not limited to, the following drugs for treating obesity: peptide YY or its analogues, neuropeptide Y receptor type 2 agonists, melanocortin receptor 4 agonists, pancreatic amyloid peptides, GIPR agonists, phosphodiesterase, AMPK, neuropeptide Y5 receptor antagonists, GPR40 agonists, GLP-1 / GIP dual receptor agonists, naltrexone / bupropion, lorcaserin, phentermine / topiramate, orlistat, or GLP-1 peptide analogues.

[0109] Drugs or active ingredients that can be used in combination with the compounds described in this invention include, but are not limited to, the following drugs for treating NASH: FXR receptor agonists, PPARα / δ agonists, fibroblast growth factor 19 / 21 analogs, thyroid hormone receptor β agonists, SGLT1 and / or SGLT2 inhibitors, acetyl-CoA carboxylase inhibitors, chemokine receptor-2 / 5 inhibitors, anti-apoptotic signal-regulated kinase 1 inhibitors, ATP-binding transporter 1 agonists, 5-lipoxygenase inhibitors, or vascular adhesion protein 1 inhibitors.

[0110] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 20–500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skills of a skilled physician.

[0111] Preparation method

[0112] The compounds in this invention can be prepared through various steps and synthetic routes. The following synthetic schemes are representative general synthetic methods. The specific synthetic routes in conjunction with the examples constitute the synthetic methods of this invention, but are not limited to the following methods:

[0113]

[0114]

[0115] Option 3

[0116]

[0117] For the synthesis of the aforementioned compound fragment Q2, Scheme 4 refers to the following two synthetic schemes:

[0118] Method 1

[0119] Method 2

[0120] Option 5

[0121]

[0122] Option Six

[0123]

[0124] Option 7

[0125] The present invention will be further illustrated by the following examples. It should be particularly noted that these examples are for illustrative purposes only and are not intended to limit the invention in any way. Unless otherwise specified, the packing material used for column chromatography is silica gel. Experimental methods in the following examples, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer.

[0126] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be used in this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0127] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions (such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989)) or as recommended by the manufacturer. All parameters and other descriptions in the examples, unless otherwise stated, are based on mass. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0128] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0129] Example 1: (S)-2-((4-(6-((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzi[d]imidazolium-6-carboxylic acid

[0130]

[0131]

[0132] 1: 3-Bromopropene; 2: NaIO4, K2OsO4·9H2O; 3: tert-butoxyformylhydrazine, NaBH3CN, sodium triacetoxyborohydride; 4: 4M hydrogen chloride (1,4-dioxane solution); 5: trimethyl orthoformate, glacial acetic acid; 6: Pd2(dba)3, Ruphos, Cs2CO3, toluene, 120℃; 7: 4M hydrochloric acid, DCM / CH3OH = 1:1; 8: K2CO3, KI, acetonitrile, 80℃; 9: NaOH, THF / CH3OH / H2O = 3:3:1

[0133] The intermediate compound (0105-1) was synthesized according to the method described for intermediate 0116-3 in CN202110839013.8. The synthetic route is as follows:

[0134]

[0135] a, K₂CO₃, DMF; b, MeOH, Pd / CH₂; c, 2-chloro-1,1,1-trimethoxyethane, TsOH·H₂OTHF

[0136] Step 1: 2 g (7.32 mmol) of 6-bromo-2-tert-butyloxycarbonylaminopyridine was dissolved in 30 mL of N,N-dimethylformamide. 3.04 g (21.97 mmol) of anhydrous potassium carbonate and 1.9 mL (21.97 mmol) of 3-bromopropene were added. The mixture was reacted in an oil bath at 70 °C for approximately 3 hours, monitored by LC-LC. After the reaction, the filtrate was extracted three times with ethyl acetate, washed three times with saturated sodium chloride brine, dried over magnesium sulfate, and filtered. The filtrate was mixed with 100-200 mesh silica gel and subjected to Flash-LC column chromatography to obtain approximately 2.17 g of tert-butylallyl (6-bromopyridin-2-yl)carbamate oil (0203-1). LC-LC (ESI): m / z = 312.1 (M+H) + .

[0137] Step 2: 2.17 g (6.95 mmol) tert-butylallyl(6-bromopyridin-2-yl)carbamate (0203-1) was dissolved in 50 ml of a 1:1 mixture of tetrahydrofuran and water. 25.6 mg (0.0695 mmol) of potassium (VI) osmium tetroxide dihydrate and 7.44 g (34.75 mmol) of sodium periodate were added. The reaction was carried out at room temperature for about 1 hour, during which a large amount of white solid precipitated. The reaction was monitored by TLC. After the reaction was completed, about 100 ml of saturated sodium thiosulfate aqueous solution was added. After stirring for about 10 minutes, the mixture was extracted three times with ethyl acetate. The organic layer was dried with magnesium sulfate and filtered. The filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain about 1.126 g of tert-butyl(6-bromopyridin-2-yl)(2-oxyethyl)carbamate oil (0204-1). LCMS(ESI): m / z = 314.03(M+H) + .

[0138] Step 3: 1.126 g (3.59 mmol) of tert-butyl(6-bromopyridin-2-yl)(2-oxyethyl)carbamate (0204-1) was dissolved in 30 mL of 1,2-dichloroethane. 0.95 g (7.2 mmol) of tert-butoxyformylhydrazine was added, and the reaction was carried out at room temperature for approximately 1 hour, monitored by TLC. After the reaction was complete, 2.28 g (10.77 mmol) of sodium triacetoxyborohydride and 677 mg (10.77 mmol) of sodium cyanoborohydride were added, and the reaction was carried out for approximately 3 hours, monitored by LCMS. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate aqueous solution, extracted twice with dichloromethane, the organic layer was dried over magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 1.12 g of tert-butyl2-(2-((6-bromopyridin-2-yl)(tert-butoxycarbonyl)). Amino(ethyl)hydrazine-1-carboxylic acid ester oil (0205-1). LCMS (ESI): m / z = 430.12 (M+H)+ .

[0139] Step 4: 1.12 g (2.592 mmol) of tert-butyl-2-(2-((6-bromopyridin-2-yl)(tert-butoxycarbonyl)amino)ethyl)hydrazine-1-carboxylic acid ester (0205-1) was dissolved in 24 mL of dichloromethane. 12 mL of a 4M solution of 1,4-dioxane with hydrogen chloride was added. The reaction was allowed to proceed for approximately 5 hours, resulting in the precipitation of a large amount of white solid. The reaction was monitored by LCMS. After the reaction was complete, the solvent was directly evaporated to obtain the hydrochloride salt of 6-bromo-N-(2-hydrazylethyl)pyridin-2-amine (0206-1), which can proceed directly to the next step without purification. LCMS (ESI): m / z = 230.02 (M+H) + .

[0140] Step 5: Dissolve the hydrochloride salt of 6-bromo-N-(2-hydrazinoethyl)pyridine-2-amine (0206-1) from the previous step in 24 ml of glacial acetic acid, add 12 ml of trimethyl orthoformate, and react overnight at 100°C under nitrogen protection. Evaporate the solvent using a rotary evaporator, add saturated sodium bicarbonate aqueous solution, and extract three times with ethyl acetate. Dry the organic layer with magnesium sulfate, filter, and mix the filtrate with 100-200 mesh silica gel. Analyze the filtrate by Flash-LC column chromatography to obtain approximately 560 mg (2.09 mmol) of 4-(6-bromopyridine-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-carbonaldehyde (0207-1). LCMS (ESI): m / z = 268.0 (M+H) + .

[0141] Step 6: A mixture of 560 mg (2.09 mmol) of 4-(6-bromopyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-formaldehyde (0207-1), 402 mg (2.51 mmol) of (4-chloro-2-fluorophenyl)methanol (0301-1), 96 mg (0.105 mmol) of tris(dibenzylacetone)dipalladium, 98 mg (0.21 mmol) of 2-bicyclohexylphosphine-2',6'-diisopropoxybiphenyl, and 1.7 g (5.23 mmol) of cesium carbonate in toluene (80 ml) was placed in a sealed tube, purged with nitrogen for 5 minutes, and reacted overnight at 120 °C. After cooling, the solution was filtered, mixed with 100-200 mesh silica gel powder, and subjected to Flash-LC column chromatography to obtain approximately 420 mg of [the solution / container]. 4-(6-((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-carbonaldehyde (0208-1). LCMS (ESI): m / z = 348.1 (M+H) + .

[0142] Step 7: 400 mg (1.15 mmol) of 4-(6-((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-carbonaldehyde was dissolved in 20 ml of a 1:1 (v / v) mixture of dichloromethane and methanol. Under ice bath conditions, 1.2 ml (approximately 4.8 mmol) of 4M hydrochloric acid aqueous solution was added, and the mixture was reacted at room temperature overnight. After overnight reaction, a large amount of saturated sodium bicarbonate aqueous solution was added, and the mixture was extracted with dichloromethane. The organic layer was dried over magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel powder. Flash-LC column chromatography was used to obtain approximately 380 mg (1.1 mmol) of 4-(6-((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-1,4,5,6-tetrahydro-1,2,4-triazine (0209-1). LCMS(ESI): m / z = 320.1(M+H) + .

[0143] Step 8: 380 mg (1.1 mmol) of 4-(6-(((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-1,4,5,6-tetrahydro-1,2,4-triazine, 456 mg (3.3 mmol) potassium carbonate, 33.2 mg (0.22 mmol) potassium iodide, and 388 mg (1.32 mmol) (S)-2-(chloromethyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester were added to 30 mL of acetonitrile. The reaction was carried out at 80 °C for approximately 3 hours, monitored by LC-MS. After the reaction was complete, the mixture was filtered, and the filtrate was mixed with 100-200 mesh silica gel powder. Flash-LC column chromatography was used to obtain approximately 420 mg of (S)-2-((4-(6-(((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6- Dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester (0106-1). LCMS(ESI): m / z = 578.2 (M+H) + .

[0144] Step 9: 400 mg (0.69 mmol) of (S)-2-((4-(6-(((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester was dissolved in 21 ml of a 3:3:1 mixture of tetrahydrofuran / methanol / water. Under ice bath conditions, 83 mg (2.07 mmol) of sodium hydroxide was added, and the reaction was carried out at room temperature for approximately 8 hours. After the reaction was complete, 30 ml of water was added, and the organic solvent was evaporated to dryness using a rotary evaporator, leaving an aqueous solution. The pH was carefully adjusted to 7 with 1 M hydrochloric acid, and sodium chloride was added until supersaturated. The solution was then treated with ethyl acetate / methanol at a ratio of 10:1. Extraction was performed multiple times with a mixed solvent until the aqueous phase showed almost no fluorescence absorption when spotted under UV light using a capillary tube. The organic layer was dried with magnesium sulfate, filtered, and mixed with 100-200 mesh silica gel powder. Flash-LC column chromatography yielded approximately 320 mg of the final product (S)-2-((4-(6-((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-1). LCMS (ESI): m / z = 564.2 (M+H) + . 1 H NMR (500MHz, DMSO-d6) δ8.14(d,J=1.4Hz,1H),7.96(s,1H), 7.82(dd,J=8.3,1.4Hz,1H),7.66(t,J=8.0Hz,1H),7.58–7.46(m,3H),7.31(dd,J= 8.2,2.1Hz,1H),6.54(d,J=8.0Hz,1H),6.42(d,J=7.9Hz,1H),5.37(s,2H),5.07(qd,J =7.1,3.1Hz,1H),4.77(dd,J=15.4,7.1Hz,1H),4.65(dd,J=15.3,3.1Hz,1H),4.48(dt, J=13.4,3.6Hz,2H),4.40–4.31(m,2H),3.77(t,J=5.2Hz,2H),3.09(hept,J=5.9,5.2 Hz,2H),2.73–2.63(m,1H),2.46–2.36(m,1H).

[0145] Example 2, (S)-2-(4-(6-((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazol-6-carboxylic acid.

[0146]

[0147] The experimental procedures were as described in Example 1, except that the right-hand intermediate compound (0105-2) was prepared from the starting compound 3,5-difluoro-4-nitrobenzonitrile (0101-2) using the general synthetic route scheme 1. The specific experimental steps are as follows:

[0148]

[0149] 1: CH3OH, SOCl2; 2: ((2S)-oxetane-2-yl)methylamine, K2CO3, DMF, 80℃; 3: Pd / C, H2; 4: p-TsOH (cat.), 2-chloro-1,1,1-trimethoxyethane, acetonitrile

[0150] Step 1: Add 2.4 ml of SOCl2 to 10 ml of anhydrous methanol under an ice-salt bath, then add 600 mg (3.26 mmol) of 3,5-difluoro-4-nitrobenzenenitrile (0101-2). React overnight at room temperature, then heat to 50 °C, stir for half an hour, then heat to 80 °C and reflux for 2 hours. Monitor the reaction by TLC. When the reaction is complete, quench the reaction with saturated NaHCO3 aqueous solution. Extract twice with ethyl acetate. Dry the organic layer with anhydrous magnesium sulfate, filter, mix with 100-200 mesh silica gel powder, and precipitate by Flash-LC column chromatography to obtain approximately 630 mg of methyl 3,5-difluoro-4-nitrobenzene (0102-2).

[0151] Step 2: 630 mg (2.90 mmol) of methyl 3,5-difluoro-4-nitrobenzoate (0102-2) was dissolved in 20 ml of N,N-dimethylformamide, and 668 mg (4.83 mmol) of anhydrous potassium carbonate and 211 mg (2.42 mmol) of (S)-oxetane-2-methylamine were added. The reaction was carried out at 80 °C and monitored by TLC. After the reaction was completed, the mixture was filtered, extracted twice with ethyl acetate, washed three times with saturated brine, dried over anhydrous magnesium sulfate, filtered, and mixed with 100-200 mesh silica gel powder. The mixture was then subjected to Flash-LC column chromatography to obtain approximately 630 mg of (S)-3-fluoro-4-nitro-5-((oxetane-2-ylmethyl)amino)benzoate (0103-2).

[0152] Step 3: Dissolve 630 mg of (S)-2-fluoro-4-nitro-5-((oxetane-2-ylmethyl)amino)benzoate (0103-2) in 40 ml of ethyl acetate, add 60 mg of 10% palladium on carbon (55% aqueous), displace hydrogen gas, and react for approximately 2 hours. After the reaction is complete, filter the palladium on carbon, evaporate the solvent to dryness, and obtain approximately 550 mg of (S)-3-fluoro-4-amino-5-((oxetane-2-ylmethyl)amino)benzoate (0104-2). LCMS (ESI): m / z = 254.11 (M+H) + .

[0153] Step 4: Dissolve 550 g (2.16 mmol) of (S)-3-fluoro-4-amino-5-((oxetane-2-ylmethyl)amino)methyl benzoate (0104-2) in 40 ml of acetonitrile. Add 665 mg (4.32 mmol, 580 μl) of 2-chloro-1,1,1-trimethoxyethane and 82 mg (0.43 mmol) of p-toluenesulfonic acid monohydrate. React at 60 °C, monitored by TLC. After the reaction is complete, directly mix with 100-200 mesh silica gel powder and perform Flash-LC column chromatography to obtain approximately 600 mg of (S)-2-(chloromethyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester (0105-2). LCMS (ESI): m / z = 312.1 (M+H) + .

[0154] The subsequent reaction followed the procedure of Example 1, replacing intermediate (0105-1) with intermediate (0105-2) obtained by the above method, with the remaining steps identical, ultimately yielding (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-2). LCMS (ESI): m / z = 573.2 (M+H) + . 1H NMR (500MHz, DMSO-d6) δ7.96(s,1H),7.89(s,1H),7.66(t,J=8.0Hz,1H),7.55(t,J=8.2Hz,1H),7.51–7.44(m, 2H),7.31(dd,J=8.2,2.1Hz,1H),6.54(d,J=8.0Hz,1H),6.42(d,J=7.9Hz,1H),5.37 (s,2H),5.07(qd,J=7.0,3.1Hz,1H),4.76(dd,J=15.4,7.0Hz,1H),4.64(dd,J=15.3, 3.1Hz,1H),4.52–4.44(m,2H),4.40–4.31(m,2H),3.78(t,J=5.1Hz,2H),3.11(hept,J=6.2,5.5Hz,2H),2.74–2.65(m,1H),2.45–2.36(m,1H).

[0155] Example 3, (S)-2-((4-(6-((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-5-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-3).

[0156]

[0157] The experimental procedures were as described in Example 1, except that the right-side intermediate compound (0105-3) was prepared from the starting compound 2,5-difluoro-4-nitrobenzoic acid (0101-3) using the general synthetic route scheme 1. The specific experimental steps are as follows:

[0158]

[0159] 1: CH3OH, H2SO 42 ; 2: ((2S)-oxetane-2-yl)methylamine, K2CO3, DMF, 80℃; 3: Pd / C, H2; 4: p-TsOH (cat.), 2-chloro-1,1,1-trimethoxyethane, acetonitrile

[0160] Step 1: 1 g (4.93 mmol) of 2,5-difluoro-4-nitrobenzoic acid (0101-3) was dissolved in 20 ml of methanol, and 0.5 ml of concentrated sulfuric acid was added dropwise. The mixture was refluxed and reacted for about 5 hours under TLC monitoring. After the reaction was completed, most of the solvent was evaporated, and the mixture was carefully poured into a saturated sodium bicarbonate aqueous solution. The mixture was extracted three times with ethyl acetate, and the organic layer was dried with magnesium sulfate. The solvent was then filtered and evaporated to obtain about 1.06 g of methyl 2,5-difluoro-4-nitrobenzoate (0102-3).

[0161] Step 2: 1.06 g (4.88 mmol) of methyl 2,5-difluoro-4-nitrobenzoate (0102-3) was dissolved in 30 ml of acetonitrile, and 878 mg (6.35 mmol) of anhydrous potassium carbonate and 426 mg (4.88 mmol) of (S)-oxetane-2-methylamine were added. The reaction was carried out at 80 °C and monitored by TLC. After the reaction was completed, the mixture was filtered and mixed with 100-200 mesh silica gel powder. The sample was then subjected to Flash-LC column chromatography to obtain approximately 1.16 g of (S)-2-fluoro-4-nitro-5-((oxetane-2-ylmethyl)amino)benzoate (0103-3).

[0162] Step 3: Dissolve 1.16 g (4.08 mmol) of (S)-2-fluoro-4-nitro-5-((oxetane-2-ylmethyl)amino)methyl benzoate (0103-3) in 40 mL of ethyl acetate, add 116 mg of 10% palladium on carbon (55% aqueous), displace hydrogen gas, and react overnight. After the reaction is complete, filter the palladium on carbon, evaporate the solvent to dryness, and obtain approximately 1.02 g of (S)-2-fluoro-4-amino-5-((oxetane-2-ylmethyl)amino)methyl benzoate (0104-3). LCMS (ESI): m / z = 254.11 (M+H) + .

[0163] Step 4: Dissolve 1.02 g (4.01 mmol) of (S)-2-fluoro-4-amino-5-((oxetane-2-ylmethyl)amino)methyl benzoate (0104-3) in 40 ml of acetonitrile, add 927 mg (6.02 mmol, 808 μl) of 2-chloro-1,1,1-trimethoxyethane and 76 mg (0.4 mmol) of p-toluenesulfonic acid monohydrate, and react at 60 °C. Monitor the reaction by TLC. After the reaction is complete, mix the sample directly with 100-200 mesh silica gel powder and perform Flash-LC column chromatography to obtain approximately 1.12 g of (S)-2-(chloromethyl)-5-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester (0105-3). LCMS (ESI): m / z = 312.1 (M+H) + .

[0164] The subsequent reaction followed the procedure of Example 2, except that intermediate (0105-1) was replaced with intermediate (0105-3), while the remaining steps were identical, ultimately yielding (S)-2-((4-(6-(((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-5-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-3). LCMS (ESI): m / z = 582.2 (M+H) + . 1 H NMR (500MHz, DMSO-d6) δ8.22(d,J=6.4Hz, 1H),7.95(s,1H),7.66(t,J=8.0Hz,1H),7.54(t,J=8.2Hz,1H),7.51–7.45(m,2H), 7.31(dd,J=8.2,2.1Hz,1H),6.55(d,J=8.0Hz,1H),6.42(d,J=8.0Hz,1H),5.36(s, 2H),5.05(qd,J=7.1,2.7Hz,1H),4.83(dd,J=15.4,7.3Hz,1H),4.69(dd,J=15.4,2.7 Hz,1H),4.48(td,J=9.0,6.6Hz,2H),4.40–4.32(m,2H),3.78(t,J=5.3Hz,2H),3.20–3.07(m,2H),2.73–2.63(m,1H),2.44–2.34(m,1H).

[0165] Example 4, 2-((4-(6-((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((1-ethyl-1H-imidazol-5-yl)methyl)-1Hbenzi[d]imidazol-6-carboxylic acid (0107-4).

[0166]

[0167] The synthesis method is described in Example 1, wherein the intermediate on the right is replaced by compound (0105'-1), and the intermediate compound (0105'-1) is prepared by the following synthetic route.

[0168]

[0169] 1. EtOTf, dichloromethane, rt; 2. Water / acetone = 1:2, rt; 3. NH2OH hydrochloride, Na2CO3, H2O, rt to 70℃; 4. Pd / C, H2; 5. K2CO3, acetonitrile, 80℃; 6. Pd / C, H2, glacial acetic acid; 7. p-TsOH (cat.), acetonitrile, 60℃.

[0170] Step 1: 5 g (14.78 mmol) of 1-triphenylmethylimidazolium-4-carboxaldehyde was dissolved in 50 ml of ultra-dry dichloromethane. The solution was purged with nitrogen, and 3.95 g (22.17 mmol) of ethyl trifluoromethanesulfonate dissolved in 20 ml of ultra-dry dichloromethane was added dropwise at room temperature. The reaction was allowed to proceed overnight. Most of the dichloromethane was evaporated, and a large amount of cyclohexane was slowly added while stirring vigorously. A large amount of white solid precipitated from the solution. The solid was filtered and dried in a vacuum drying oven to obtain 6.2 g of the compound 3-ethyl-4-formyl-1-triphenylmethyl-1H-imidazolium-3-trifluoromethanesulfonate quaternary ammonium salt.

[0171] Step 2: Dissolve 6.2g of the obtained compound 3-ethyl-4-formyl-1-triphenylmethyl-1H-imidazolium-3-trifluoromethanesulfonic acid quaternary ammonium salt in 120ml of a mixed solvent of H2O / acetone = 1 / 2. React at room temperature and monitor with TLC. After the reaction is complete, add saturated sodium chloride aqueous solution and extract three times with ethyl acetate. Dry the organic layer with anhydrous magnesium sulfate, filter, mix the filtrate with 100-200 mesh silica gel, and precipitate by Flash-LC column chromatography to obtain approximately 1.3g of 1-ethyl-1H-imidazolium-5-carboxaldehyde.

[0172] Step 3: Add 2.19 g (31.45 mmol) of NH2OH hydrochloride and 3.33 g (31.45 mmol) of sodium carbonate to 50 ml of water and stir for about 15 minutes. Then add 1.3 g (10.48 mmol) of 1-ethyl-1H-imidazol-5-carboxaldehyde obtained in the previous step, and react at 70 °C for 3-4 hours. During the reaction, from the dissolved state to the appearance of a large amount of precipitate, the reaction was monitored by TLC until it was complete. After cooling to room temperature, filter the solid and dry it under vacuum. No further purification is required to obtain 1.23 g of 1-ethyl-1H-imidazol-5-carboxaldehyde oxime.

[0173] Step 4: Dissolve the product from the previous step in 20 ml of glacial acetic acid, add 240 mg of Pd / C (10% Pd content, 55% water content), and react with hydrogen gas. After about 5 hours, monitor the reaction with TLC until it ends. Filter the Pd / C and evaporate the glacial acetic acid directly to obtain about 1.1 g of (1-ethyl-1H-imidazol-5-yl)methylamine (1102), which is then directly added to the next step.

[0174] Step 5: Dissolve 1.1 g (8.8 mmol) (1-ethyl-1H-imidazol-5-yl)methylamine (1102) in 20 ml N,N-dimethylformamide, add 3.65 g (26.4 mmol) anhydrous potassium carbonate and 1.75 g (8.8 mmol) methyl 3-fluoro-4-nitrobenzene (0102-1), react at 80 °C for about 2 hours, monitor by TLC. After the reaction is complete, filter out the insoluble salt, pour the filtrate into saturated brine, extract with ethyl acetate 3 times, and then wash with saturated brine 3 times. Dry the organic layer with anhydrous magnesium sulfate, filter, mix with 100-200 mesh silica gel, and precipitate by Flash-LC column chromatography to obtain about 1.73 g of methyl 3-((1-ethyl-1H-imidazol-5-yl)methyl)amino)-4-nitrobenzene (0103-1).

[0175] Step 6: Dissolve the product from the previous step in 50 ml of ethyl acetate, add 150 mg of Pd / C (10% Pd content, 55% water content), replace with hydrogen, react overnight, monitor the reaction by TLC, after the reaction is complete, filter Pd / C, evaporate the solvent to dryness, and obtain 1.48 g of methyl 4-amino-3-((1-ethyl-1H-imidazol-5-yl)methylamino)benzoate (0104'-1).

[0176] Step 7: Dissolve 1.48 g (5.4 mmol) of methyl 4-amino-3-((1-ethyl-1H-imidazol-5-yl)methylamino)benzoate (0104'-1) in 40 ml of acetonitrile, add 1.247 g (8.1 mmol, 1.09 ml) of 2-chloro-1,1,1-trimethoxyethane and 103 mg (0.54 mmol) of p-toluenesulfonic acid monohydrate, and react at 60 °C. Monitor the reaction by TLC. After the reaction is complete, directly mix with 100-200 mesh silica gel and precipitate by Flash-LC column chromatography to obtain approximately 1.29 g of methyl 2-(chloromethyl)-1-(1-ethyl-1H-imidazol-5-yl)methyl)-1Hbenzo[d]imidazolium-6-carboxylic acid (0105'-1). LCMS (ESI): m / z = 332.1 (M+H) + .

[0177] The subsequent reaction followed the procedure of Example 1, except that intermediate (0105-1) was replaced with intermediate (0105-1), and the remaining steps were identical, ultimately yielding 2-((4-(6-(((4-chloro-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((1-ethyl-1H-imidazol-5-yl)methyl)-1Hbenzo[d]imidazol-6-carboxylic acid (0107-4). LCMS (ESI): m / z = 602.2 (M+H) + . 1H NMR (500MHz, DMSO-d6) δ8.12(d,J=1.6Hz,1H),7.94(s,1H), 7.84(dd,J=8.4,1.6Hz,1H),7.70(d,J=8.4Hz,1H),7.68–7.63(m,2H),7.54(t,J=8.2 Hz,1H),7.49(dd,J=9.9,2.1Hz,1H),7.32(dd,J=8.2,2.1Hz,1H),6.52(d,J=8.0Hz,1H ),6.48(s,1H),6.43(d,J=8.0Hz,1H),5.76(s,2H),5.36(s,2H),4.38(s,2H),3.96(q, J=7.2Hz,2H),3.66(t,J=5.1Hz,2H),3.05(t,J=5.1Hz,2H),1.13(t,J=7.2Hz,3H).

[0178] Example 5, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-5).

[0179]

[0180] The experimental procedure was the same as in Example 1, using the same intermediate compound (0207-1), but replacing (4-chloro-2-fluorophenyl)methanol (0301-1) with (4-cyano-2-fluorophenyl)methanol (0301-2). The remaining steps were identical, ultimately yielding (S)-2-((4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-5). LCMS (ESI): m / z = 555.2 (M+H) + . 1HNMR(500MHz,DMSO-d6)δ12.80(s,1H),8.29(d,J=1.5Hz, 1H),7.94–7.88(m,2H),7.83(dd,J=8.5,1.6Hz,1H),7.75–7.64(m,4H),6.56(d,J=8 .0Hz,1H),6.46(d,J=8.0Hz,1H),5.46(s,2H),5.07(qd,J=7.1,2.8Hz,1H),4.83(dd, J=15.4,7.2Hz,1H),4.70(dd,J=15.4,2.8Hz,1H),4.54–4.44(m,2H),4.41–4.31(m, 2H), 3.76 (t, J=5.2Hz, 2H), 3.12 (qt, J=11.2, 5.0Hz, 2H), 2.69 (ddt, J=14.2, 11.4, 7.2 Hz, 1H), 2.40 (ddt, J=11.2, 9.1, 6.9Hz, 1H).

[0181] Example 6, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-6).

[0182]

[0183] The synthesis route is the same as in Examples 2 and 5. LCMS(ESI): m / z = 573.2(M+H) + . 1 H NMR(500MHz,DMSO-d6)δ8.07(s,1H),7.91(d,J=9.0Hz,2H),7.75–7.64(m,3H),7.51(d,J=11.6Hz,1H),6.56(d,J=8.0Hz,1H),6.4 6(d,J=8.0Hz,1H),5.47(s,2H),5.06(dt,J=9.7,4.9Hz,1H),4.82(dd,J=15.4,7.1Hz,1H),4.70(dd,J=15.4,2.9Hz,1H),4.49(td, J=9.8,4.6Hz,2H),4.42–4.31(m,2H),3.77(t,J=5.2Hz,2H),3.12(qt,J=11.2,5.0Hz,2H),2.68(dt,J=16.1,7.8Hz,1H),2.40(p,J=7.4Hz,1H).

[0184] Example 7, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-5-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-7).

[0185]

[0186] The experimental procedures were the same as in Examples 3 and 5. LCMS(ESI): m / z = 573.2(M+H) + . 1 H NMR(500MHz,DMSO-d6)δ8.19(d,J=6.4Hz,1H),7.93–7.88(m,2H),7.75–7.63(m,3H),7.48 (d,J=11.6Hz,1H),6.56(d,J=7.9Hz,1H),6.46(d,J=8.0Hz,1H),5.46(s,2H),5.05 (qd,J=7.2,2.8Hz,1H),4.82(dd,J=15.4,7.3Hz,1H),4.68(dd,J=15.3,2.8Hz,1H), 4.51–4.44(m,2H),4.39–4.31(m,2H),3.76(t,J=5.3Hz,2H),3.20–3.02(m,2H), 2.73–2.63(m,1H),2.39(ddt,J=11.1,9.0,7.0Hz,1H).

[0187] Example 8, 2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((1-ethyl-1H-imidazol-5-yl)methyl)-1Hbenzo[d]imidazol-6-carboxylic acid (0107-8).

[0188]

[0189] The experimental procedures were as described in Examples 4 and 5. LCMS(ESI): m / z = 593.2(M+H) + . 1H NMR(500MHz,DMSO-d6)δ8.11(d,J=1.6Hz,1H),7.93–7.88(m,2H),7.84(dd,J=8.5,1.6Hz, 1H),7.74–7.64(m,5H),6.53(d,J=8.0Hz,1H),6.49–6.44(m,2H),5.76(s,2H),5.46 (s,2H),4.38(s,2H),3.95(q,J=7.2Hz,2H),3.64(t,J=5.2Hz,2H),3.05(t,J=5.2Hz, 2H),1.13(t,J=7.2Hz,3H).

[0190] Example 9, (S)-2-((4-(6-((4-cyano-2-methoxybenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-9).

[0191]

[0192] The synthetic route was the same as in Example 1, except that the starting compound 4-(hydroxymethyl)-3-methoxybenzonitrile (0301-3) replaced compound (0301-1). LCMS (ESI): m / z = 567.2 (M+H) + . 1 H NMR (500MHz, DMSO-d6) δ8.28(s,1H),7.83(s,1H),7.67(m,2H),7.54–7.46(m,3H),7.43(dd,J=7.7,1.5Hz, 1H),6.53(d,J=8.0Hz,1H),6.47(d,J=8.0Hz,1H),5.35(s,2H),5.09(qd,J=7.0,2.9 Hz,1H),4.83(dd,J=15.4,7.1Hz,1H),4.70(dd,J=15.3,2.9Hz,1H),4.48(td,J=8.7, 6.4Hz,2H),4.39–4.31(m,2H),3.88(s,3H),3.73(t,J=5.2Hz,2H),3.13(dp,J=16.9, 6.2,5.5Hz,2H),2.70–2.64(m,1H),2.39–2.32(m,1H).

[0193] Example 10, (S)-2-((4-(6-((4-cyano-2-methoxybenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1Hbenzi[d]imidazolium-6-carboxylic acid (0107-10).

[0194]

[0195] The synthesis route is the same as in Examples 2 and 9. LCMS(ESI): m / z = 585.2(M+H) + . 1 H NMR (500MHz, DMSO-d6) δ8.06 (s, 1H), 7.87 (s, 1H), 7.67 (t, J = 8.0Hz, 1H), 7.54–7.48 (m, 3H), 7.42(dd,J=7.7,1.5Hz,1H),6.53(d,J=8.0Hz,1H),6.47(d,J=8.0Hz,1H),5.37(s, 2H),5.07(qd,J=7.0,2.9Hz,1H),4.83(dd,J=15.4,7.1Hz,1H),4.70(dd,J=15.3,2.9 Hz,1H),4.48(td,J=8.7,6.4Hz,2H),4.41–4.31(m,2H),3.89(s,3H),3.75(t,J=5.2 Hz,2H),3.12(dp,J=16.9,6.2,5.5Hz,2H),2.74–2.65(m,1H),2.43–2.36(m,1H).

[0196] Example 11, (S)-2-((4-(6-((4-(difluoromethyl)-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-11).

[0197]

[0198] The synthetic route was the same as in Example 1, except that the starting compound (0301-1) was replaced with (4-(difluoromethyl)-2-fluorophenyl)methanol (0301-4). LCMS (ESI): m / z = 580.2 (M+H) + . 1H NMR(500MHz, DMSO-d6)δ 8.06(d,J=1.3Hz,1H),7.94(s,1H),7.81(dd,J=8.3,1.4Hz,1H),7.66(td,J=7.8,2.0 Hz,2H),7.48(d,J=10.3Hz,1H),7.44(d,J=8.2Hz,2H),7.06(t,J=55.6Hz,1H),6.54 (d,J=8.0Hz,1H),6.44(d,J=7.9Hz,1H),5.44(s,2H),5.07(qd,J=6.9,3.2Hz,1H), 4.73(dd,J=15.4,6.9Hz,1H),4.62(dd,J=15.3,3.2Hz,1H),4.52–4.44(m,2H),4.38– 4.31(m,2H),3.76(t,J=5.2Hz,2H),3.09(q,J=4.7Hz,2H),2.69(ddt,J=14.0,11.1,6.9Hz,1H),2.42(ddt,J=11.2,9.0,7.0Hz,1H).

[0199] Example 12, (S)-2-((4-(6-((4-(difluoromethyl)-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-12).

[0200]

[0201] The synthesis route is the same as in Examples 2 and 11. LCMS(ESI): m / z = 598.2(M+H)+. 1H NMR(500MHz,DMSO-d6)δ7.93(d,J=14.3Hz,2H),7.66(td,J=7.8,3.3Hz,2H),7.49(dd ,J=11.2,7.0Hz,2H),7.43(d,J=7.9Hz,1H),7.05(t,J=55.6Hz,1H),6.55(d,J=8.0Hz, 1H),6.44(d,J=7.9Hz,1H),5.44(s,2H),5.06(tt,J=7.1,4.3Hz,1H),4.76(dd,J=15.4, 7.0Hz,1H),4.64(dd,J=15.4,3.1Hz,1H),4.52–4.44(m,2H),4.40–4.31(m,2H),3.77 (t,J=5.1Hz,2H),3.10(hept,J=5.8,5.0Hz,2H),2.68(dq,J=11.2,7.7Hz,1H),2.45– 2.35(m,1H).

[0202] Example 13, (S)-2-((4-(6-((4-(trifluoromethyl)-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-13).

[0203]

[0204] The synthetic route was the same as in Example 1, except that the starting compound (0301-1) was replaced with (4-(trifluoromethyl)-2-fluorophenyl)methanol (0301-5). LCMS (ESI): m / z = 598.2 (M+H) + . 1H NMR(500MHz, DMSO-d6)δ 8.57(d,J=1.6Hz,1H),8.21(s,1H),8.10(dd,J=8.5,1.6Hz,1H),8.02(dd,J=8.5,5.8 Hz,2H),7.96(t,J=8.1Hz,2H),7.90(d,J=8.0Hz,1H),6.84(d,J=7.9Hz,1H),6.74(d, J=8.1Hz,1H),5.76(s,2H),5.35(ddt,J=10.1,7.2,2.9Hz,1H),5.11(dd,J=15.4,7.1 Hz,1H),4.98(dd,J=15.5,2.9Hz,1H),4.82–4.72(m,2H),4.69–4.59(m,2H),4.05(t, J=5.1Hz,2H),3.45–3.35(m,2H),2.96(dq,J=10.8,7.7Hz,1H),2.72–2.64(m,1H).

[0205] Example 14, (S)-2-((4-(6-((4-(trifluoromethyl)-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-14).

[0206]

[0207] The synthetic route is the same as in Examples 2 and 13. LCMS(ESI): m / z = 616.2(M+H) + . 1H NMR (500MHz, DMSO-d6) δ8.15(s,1H),7.93(s,1H),7.73(d,J=8.7Hz,2H),7.68(t,J=8.0Hz, 1H),7.64–7.59(m,1H),7.55(d,J=11.3Hz,1H),6.57(d,J=8.1Hz,1H),6.46(d,J= 8.0Hz,1H),5.48(s,2H),5.07(qd,J=7.0,2.7Hz,1H),4.85(dd,J=15.5,7.1Hz,1H), 4.72(d,J=15.1Hz,1H),4.54–4.44(m,2H),4.42–4.31(m,2H),3.78(t,J=5.2Hz,2H), 3.20–3.07(m,2H),2.68(dd,J=10.9,6.9Hz,1H),2.45–2.34(m,1H).

[0208] Example 15, (S)-2-((4-(6-((4-cyanobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzo[d]imidazolium-6-carboxylic acid (0107-15).

[0209]

[0210] The synthesis of compound 15 (0107-15) was performed according to Example 1.

[0211] The left-hand intermediate 7-(hydroxymethyl)benzofuran-4-onitrile (0301-6) is prepared according to the preparation of compound 0201-1 on page 21 of the specification in patent CN113480534A. The synthetic route is as follows:

[0212]

[0213] This intermediate (0301-6) was used to replace compound (0301-1), and the remaining experiments were performed according to Example 1 to prepare compound (S)-2-((4-(6-((4-cyanobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzo[d]imidazolium-6-carboxylic acid (0107-15). LCMS (ESI): m / z = 577.21 (M+H) + . 1H NMR (500MHz, DMSO-d6) δ8.33(d,J=2.3Hz,1H), 8.29(d,J=1.5Hz,1H),7.87–7.77(m,3H),7.67(dt,J=8.0,3.8Hz,2H),7.54(d,J=7.8 Hz,1H),7.22(d,J=2.2Hz,1H),6.55(d,J=8.0Hz,1H),6.47(d,J=7.9Hz,1H),5.71(s, 2H),5.06(td,J=7.2,2.8Hz,1H),4.84(dd,J=15.4,7.2Hz,1H),4.70(dd,J=15.4,2.8 Hz,1H),4.49(td,J=9.2,6.7Hz,2H),4.40–4.31(m,2H),3.72(t,J=5.2Hz,2H),3.10 (ddq,J=16.8,11.2,5.0Hz,2H),2.74–2.65(m,1H),2.40(m,1H).

[0214] Example 16, (S)-2-((4-(6-((4-cyanobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-16).

[0215]

[0216] The synthesis route is the same as in Examples 2 and 15. LCMS(ESI): m / z = 595.2(M+H) + . 1H NMR (500MHz, DMSO-d6) δ8.33(d,J=2.2Hz,1H),8.08(s,1H),7.86(s,1H),7.80(d,J=7.8Hz, 1H),7.67(t,J=8.0Hz,1H),7.57–7.49(m,2H),7.22(d,J=2.2Hz,1H),6.55(d,J=8.0 Hz, 1H), 6.47 (d, J=8.0Hz, 1H), 5.71 (s, 2H), 5.06 (tt, J=7.0, 4.2Hz, 1H), 4.83 (dd, J= 15.4,7.1Hz,1H),4.70(dd,J=15.3,2.9Hz,1H),4.52–4.44(m,2H),4.41–4.31(m,2H), 3.73(t,J=5.3Hz,2H),3.10(qt,J=11.2,4.9Hz,2H),2.74–2.63(m,1H),2.40(m,1H).

[0217] Example 17, (S)-2-((4-(6-((4-chlorobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-17).

[0218]

[0219]

[0220] The synthesis of compound 17 (0107-17) was performed according to Example 1.

[0221] The intermediate (4-chlorobenzofuran-7-yl)methanol (0301-7) was prepared according to the preparation of compound 0201-12 on page 32 of the specification in patent CN113480534A. The synthetic route is as follows:

[0222]

[0223] This intermediate (0301-7) was used to replace compound (0301-1), and the remaining experiments were performed according to Example 1 to prepare compound (S)-2-((4-(6-((4-chlorobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-17). LCMS (ESI): m / z = 586.2 (M+H) + .1 H NMR (500MHz, DMSO-d6) δ8.16 (d, J = 2.3Hz, 1H), 8.05 (t, J = 0.9Hz, 1H), 7.93 (s, 1H), 7.79 (dd, J = 8.3, 1.3Hz, 1H), 7.65 (t, J = 8.0Hz, 1H), 7.42(dd,J=8.1,6.4Hz,2H),7.36(d,J=8.0Hz,1H),7.05(d,J=2.2Hz,1H),6.53(d,J =8.1Hz,1H),6.42(d,J=8.0Hz,1H),5.61(s,2H),5.07(qd,J=6.9,3.1Hz,1H),4.75 (dd,J=15.3,7.0Hz,1H),4.62(dd,J=15.3,3.2Hz,1H),4.53–4.44(m,2H),4.39–4.32 (m,2H),3.75(t,J=5.1Hz,2H),3.08(q,J=4.7Hz,2H),2.75–2.63(m,1H),2.41(ddt,J=11.4,9.3,7.1Hz,1H).

[0224] Example 18, (S)-2-((4-(6-((4-chloro-2,3-dihydrobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-18).

[0225]

[0226] The synthetic route was the same as in Example 1, except that the starting compound (0301-1) was replaced with (4-chloro-2,3-dihydrobenzofuran-7-yl)methanol (0301-8).

[0227] The synthetic route for this intermediate (0301-8) is as follows:

[0228]

[0229] 2-Bromo-1,1-Diethoxyethane, K₂CO₃, 2. PPA, DCE, reflux, 3. Ru / CH₂, 4. LiAlH₄, 0℃

[0230] Step 1: Under nitrogen protection, 2 g (10.75 mmol) of methyl 4-chloro-2-hydroxybenzoate, 2.52 g (12.9 mmol) of 2-bromo-1,1-diethoxyethane, 2.97 g (21.5 mmol) of potassium carbonate, and 0.2 g (10 wt) of potassium iodide were added to 20 mL of a mixture of N,N-dimethylformamide and stirred overnight at 120 °C. After cooling to room temperature, the reaction was quenched with water and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to give 1.46 g of crude methyl 4-chloro-2-(2,2-diethoxyethoxy)benzoate. This compound did not require further purification and was used directly in the next step.

[0231] Step 2: Under nitrogen protection, 1.46 g (4.62 mmol) of methyl 4-chloro-2-(2,2-diethoxyethoxy)benzoate and 3.9 g (11.55 mmol) of polyphosphoric acid were added to 40 mL of 1,2-dichloroethane and refluxed overnight. After cooling to room temperature, the reaction was quenched with water (40 mL) and extracted three times with dichloromethane. The organic layer was washed with saturated sodium bicarbonate and brine, dried over anhydrous magnesium sulfate, filtered, and mixed with 100-200 mesh silica gel powder. The mixture was then subjected to Flash-LC column chromatography to obtain approximately 728 mg of methyl 4-chlorobenzofuran-7-carboxylic acid.

[0232] Step 3: Add 728 mg of methyl 4-chlorobenzofuran-7-carboxylate obtained in the previous step to 40 ml of a mixed solvent of methanol / ethyl acetate = 1:1, add approximately 73 mg of 5% Ru / C, replace with hydrogen, and react overnight to obtain approximately 720 mg of methyl 4-chloro-2,3-dihydrobenzofuran-7-carboxylate, and proceed directly to the next step.

[0233] Step 4: Dissolve 720 mg (3.4 mmol) of methyl 4-chloro-2,3-dihydrobenzofuran-7-carboxylic acid obtained in the previous step in 30 ml of ultra-dry tetrahydrofuran. Replace with nitrogen and, under ice bath conditions, add 2.13 ml (5.1 mmol) of a 2.4 M LiAlH4 tetrahydrofuran solution. The reaction is carried out for approximately 0.5 hours, monitored by TLC. After the reaction is complete, the reaction solution is slowly poured into a saturated sodium chloride aqueous solution, and 40 ml of ethyl acetate is added and stirred. The insoluble emulsified solid is then filtered through diatomaceous earth. The filtrate is extracted three times with ethyl acetate, the organic layer is dried over magnesium sulfate, filtered, and the sample is mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 560 mg of (4-chloro-2,3-dihydrobenzofuran-7-yl)methanol (0301-8).

[0234] Subsequent synthesis followed the method described in Example 1, replacing intermediate (0301-1) with intermediate (0301-8). The final product was obtained.

[0235] (S)-2-((4-(6-((4-chloro-2,3-dihydrobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazin-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-18). LCMS (ESI): m / z = 588.2 (M+H) + . 1 H NMR (500MHz, DMSO-d6) δ8.26(s,1H),7.99(s,1H), 7.83(dd,J=8.4,1.6Hz,1H),7.63(t,J=8.1Hz,2H),7.18(d,J=8.2Hz,1H),6.87(d,J= 8.3Hz,1H),6.50(d,J=8.0Hz,1H),6.39(d,J=7.9Hz,1H),5.21(s,2H),5.07(dt,J= 9.8,5.0Hz,1H),4.84(dd,J=15.4,7.1Hz,1H),4.70(dd,J=15.4,2.9Hz,1H),4.63(t, J=8.7Hz,2H),4.54–4.45(m,2H),4.42–4.32(m,2H),3.77(t,J=5.2Hz,2H),3.19(t,J =8.8Hz,2H),3.12(dq,J=11.8,6.1,5.7Hz,2H),2.69(dq,J=15.6,7.6Hz,1H),2.44– 2.36(m,1H).

[0236] Example 19, (S)-2-((4-(6-((4-chloro-2-fluorobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-19).

[0237]

[0238] Compound 19 (0107-19) was prepared according to Example 1, wherein the left-hand intermediate (4-chloro-2-fluorobenzofuran-7-yl)methanol (0301-9) was used instead of compound (0301-1), and was prepared using the following synthetic route:

[0239]

[0240] 1. Tetrahydropyran, Fe(NO3)3 9H2O, NaI; 2. LDA, -78℃, DMF; 3. 50% (wt%) NaOH (aq.), DMF; 4. Ethyl bromoacetate, K2CO3, 80℃ to 130℃; 5. NaOH; 6. SeletF, KF, 70℃, sealed tube, DCE / H2O = 2:1; 7. PPTS, MeOH, reflux.

[0241] Step 1: 760 mg (5 mmol) NaI was added to 150 ml dichloromethane, followed by 1 g (2.5 mmol) ferric nitrate nonahydrate. The solution quickly turned a pale purplish-red color. Then, 20 g (0.125 mol) of (4-chloro-2-fluorophenyl) methanol and 14.72 g (0.175 mol) tetrahydropyran were added. The reaction was slightly exothermic and lasted for about 1 hour. The reaction was monitored by TLC and then stopped. The reaction was quenched with sodium thiosulfate aqueous solution, extracted twice with dichloromethane, dried over anhydrous magnesium sulfate, filtered, mixed with 100-200 mesh silica gel, and subjected to Flash-LC column chromatography to obtain 15.7 g of 2-((4-chloro-2-fluorobenzyl)oxy)tetrahydro-2H-pyran.

[0242] Step 2: 15.7 g (64.2 mmol) of 2-((4-chloro-2-fluorobenzyl)oxy)tetrahydro-2H-pyran was dissolved in 80 ml of ultra-dry tetrahydrofuran, purged with nitrogen, cooled to -78 °C, and 35.3 ml (70.6 mmol) of 2M LDA was added dropwise. After reacting for half an hour, 5.67 ml (73.62 mmol) of ultra-dry DMF was added dropwise. After reacting for 1 hour, the reaction was monitored by TLC until the reaction was complete. The reaction solution was then slowly poured into a stirred saturated sodium chloride aqueous solution, extracted three times with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, and the filtrate was evaporated to dryness to obtain approximately 16.7 g of 6-chloro-2-fluoro-3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzaldehyde.

[0243] Step 3: 16.7 g (61.3 mmol) of 6-chloro-2-fluoro-3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzaldehyde was dissolved in 50 ml of DMF, and 50% (wt%) NaOH aqueous solution was added dropwise. The reaction was carried out at room temperature for about 1 hour. After the reaction was completed by TLC monitoring, the pH was adjusted to about 4-5 with 1M hydrochloric acid aqueous solution, saturated sodium chloride aqueous solution was added, and the mixture was extracted with ethyl acetate 3 to 4 times, washed with saturated brine more than 3 times, dried with anhydrous magnesium sulfate, filtered, mixed with 100-200 mesh silica gel, and Flash-LC column chromatography was performed to obtain about 11.8 g of 6-chloro-2-hydroxy-3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzaldehyde.

[0244] Step 4: 11.8 g (43.3 mmol) of 6-chloro-2-hydroxy-3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzaldehyde obtained in the previous step was dissolved in 70 ml of DMF. 17.95 g (0.13 mol) of K2CO3 and 14.54 ml of ethyl 2-bromoacetate were added. The reaction was carried out at 80 °C for about 3 hours. After the reaction of the starting material was completed by TLC monitoring, the temperature was raised to 130 °C and the reaction was continued for 3 to 5 hours until the new spot generated at 80 °C disappeared. The reaction was then completed. After cooling, the mixture was filtered. The filtrate was extracted and washed with an ethyl acetate / saturated sodium chloride aqueous solution system. The organic phase was dried over anhydrous magnesium sulfate, filtered, and mixed with 100-200 mesh silica gel. Flash-LC column chromatography was used to obtain approximately 7.62 g of ethyl 4-chloro-7-(((tetrahydrofuran-2H-pyran-2-yl)oxy)methyl)benzofuran-2-carboxylic acid.

[0245] Step 5: Dissolve 7.62 g (22.5 mmol) of ethyl 4-chloro-7-(((tetrahydrofuran-2H-pyran-2-yl)oxy)methyl)benzofuran-2-carboxylic acid in 70 ml of a mixed solvent of THF / CH3OH / H2O = 3:3:1. Add about 3 g of NaOH and react at room temperature for about 4 hours. After the reaction is complete as monitored by TLC, add 50 ml of pure water and evaporate the organic solvent, leaving an aqueous solution. Carefully adjust the pH to 7 with 1 M hydrochloric acid. Add a large amount of saturated sodium chloride solution to this aqueous solution, and then extract at least 4 times with a mixed solvent of EA / CH3OH = 10:1. Dry the organic phase with anhydrous magnesium sulfate, filter, and evaporate the organic solvent to obtain about 6.9 g of 4-chloro-7-(((tetrahydrofuran-2H-pyran-2-yl)oxy)methyl)benzofuran-2-carboxylic acid.

[0246] Step 6, take the 6.9 g (22.2 mmol) of 4-chloro-7-(((tetrahydrofuran-2H-pyran-2-yl)oxy)methyl obtained in the previous step. Benzofuran-2-carboxylic acid was suspended in a sealed tube containing 120 ml of a 2:1 mixture of 1,2-dichloroethane and water. Nitrogen gas was purged into the sealed tube for 3 minutes. Then, 15.73 g (44.4 mmol) of SelectF and 5.16 g (88.9 mmol) of KF were added, and nitrogen gas was purged again for 1 minute. The reaction was then carried out overnight at 70 °C. After cooling, the mixture was extracted with a DCM / NaCl (aq.) system. The organic phase was dried over anhydrous magnesium sulfate, filtered, mixed with 100-200 mesh silica gel, and subjected to Flash-LC column chromatography to obtain approximately 1.52 g of 4-chloro-2-fluoro-7-(((tetrahydrofuran-2H-pyran-2-yl)oxy)methyl)benzofuran and 480 mg of (4-chloro-2-fluorobenzofuran-7-yl)methanol (0301-9).

[0247] Step 7: Dissolve 1.52 g (3.96 mmol) of 4-chloro-2-fluoro-7-(((tetrahydrofuran-2H-pyran-2-yl)oxy)methyl)benzofuran obtained in the previous step in 40 ml of methanol, add 152 mg of p-toluenesulfonic acid pyridinium, and then react at 70 °C for about 1 hour. The reaction is monitored by TLC until it ends. After cooling, directly mix with 100-200 mesh silica gel and Flash-LC column chromatography to obtain about 752 mg of (4-chloro-2-fluorobenzofuran-7-yl)methanol (0301-9). Combine the 480 mg of compound (0301-9) obtained in step 6 to obtain a total of 1.232 g of intermediate compound (4-chloro-2-fluorobenzofuran-7-yl)methanol (0301-9).

[0248] The subsequent reaction is as described in Example 1.

[0249]

[0250]

[0251] Step 8: In a sealed tube, add 1.5 g (5.6 mmol) of compound (0207-1), 1.232 g (6.16 mmol) of compound (0301-9), and 5.02 g (15.4 mmol) of Cs₂CO₃, then add 80 ml of toluene. Purge the sealed tube with nitrogen for 3 minutes, then add 564 mg (0.616 mmol) of Pd₂(dba)₃ and 575 mg (1.232 mmol) of Ruphos. Purge with nitrogen for another minute, then react overnight at 125 °C. After cooling, filter, and directly mix the filtrate with 100-200 mesh silica gel. Flash-LC column chromatography yields approximately 847 mg of 4-(6-((4-chloro-2-fluorobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4- Triazine-1(4H)-formaldehyde (0208-19) and 780 mg of 4-(6-((2-fluorobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-formaldehyde (0208-20).

[0252] Subsequent reactions were carried out as described in Example 1, ultimately yielding the final product (S)-2-((4-(6-(((4-chloro-2-fluorobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-19). LCMS (ESI): m / z = 604.2 (M+H) + .

[0253] Example 20, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid methyl ester (0107-20).

[0254]

[0255] The synthesis of compound 20 (0107-20) was performed according to Example 5, LCMS (ESI): m / z = 569.2 (M+H). + .

[0256] Example 21, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-3-methyl-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-21).

[0257]

[0258] The synthetic route of compound 21 (0107-21) was as described in Example 5.

[0259]

[0260] The triazine intermediate (0207-2) was prepared from intermediate 0206-1 and triethyl orthoacetate according to Example 1, with the remaining steps following the same procedure. LCMS (ESI): m / z = 569.2 (M+H) + .

[0261] Example 22, 2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-6-methyl-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1Hbenzimidazole-6-carboxylic acid (0107-22).

[0262]

[0263] The synthetic route of compound 22 (0107-22) was prepared according to Example 5.

[0264]

[0265] The triazine intermediate 0203-2 was prepared by referring to the preparation of intermediate 0202-1 and 2-methyl-3-bromopropene in Example 1. Subsequent steps were similar to those in Example 1, finally yielding 2-((4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-6-methyl-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1Hbenzimidazole-6-carboxylic acid (0107-22). LCMS (ESI): m / z = 569.2 (M+H) + .

[0266] Example 23, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzi[d]imidazolium-5-carboxylic acid (0107-23)

[0267]

[0268] The synthetic route of compound 23 (0107-23) was as described in Example 5.

[0269] The right-hand intermediate (S)-2-(chloromethyl)-1-(oxetane-2-ylmethyl)-1H benzo[d]imidazolium-5-carboxylic acid methyl ester (0105-4) was prepared from the starting material 4-fluoro-3-nitrobenzoic acid according to the synthetic route of intermediate compound 0116-3 as described in CN202110839013.8. The remaining steps were the same as in Example 5. LCMS (ESI): m / z = 555.2 (M+H) + .

[0270] Example 24, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-3-(oxetane-2-ylmethyl)-3H-imidazolium[4,5-b]pyridine-5-carboxylic acid (0107-24)

[0271]

[0272] The synthetic route of compound 24 (0107-24) was prepared according to Example 5.

[0273] The right-hand intermediate (S)-2-(chloromethyl)-3-(oxetane-2-ylmethyl)-3H-imidazolium[4,5-b]pyridine-5-carboxylic acid methyl ester (0105-5) was prepared according to the synthesis of intermediate compound 0116-1 as described in CN202110839013.8. The remaining steps were the same as in Example 5. LCMS (ESI): m / z = 556.2 (M+H) + .

[0274] Example 25, 2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(2-methoxyethyl)-1H-benzo[d]imidazol-6-carboxylic acid (0107-25).

[0275]

[0276] The synthetic route of compound 25 (0107-25) was prepared according to Example 5.

[0277] The right-hand intermediate 2-(chloromethyl)-1-(2-methoxyethyl)-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester (0105-6) was synthesized according to intermediate 19 of patent WO2018109607A1, with the remaining steps the same as in Example 5. LCMS (ESI): m / z = 543.2 (M+H) + .

[0278] Example 26, 2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(tetrahydrofuran-3-yl)-1Hbenzimidazole-6-carboxylic acid (0107-26).

[0279]

[0280] The synthesis of compound 26 (0107-26) is described in Example 5.

[0281] The right-hand fragment, 2-(chloromethyl)-1-(tetrahydrofuran-3-yl)-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester (0105-7), was synthesized according to the intermediate 0116-3 in CN202110839013.8, where 3-aminotetrahydrofuran replaced (S)-oxetane-2-ylmethylamine. The synthetic route is as follows.

[0282] 1, K₂CO₃, DMF, 2, MeOH, Pd / C H₂, 3, 2-Chloro-1,1,1-Trimethoxyethane, TsOH·H₂O THF

[0283] The remaining steps are the same as in Example 5. LCMS(ESI): m / z = 555.2(M+H) + .

[0284] Example 27, 2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((1-(cyanomethyl)cyclopropyl)methyl)-4-fluoro-1H-benzo[d]imidazol-6-carboxylic acid (0107-27).

[0285]

[0286] The synthesis of compound 27 (0107-27) was performed according to Example 5, wherein the right-hand fragment 2-(chloromethyl)-1-(1-(cyanomethyl)cyclopropyl)methyl)-4-fluoro-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester (0105-8) was synthesized according to Example 2, wherein the right-hand fragment (S)-2-(chloromethyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester (0105-2) was reacted with intermediate 2-(1-(aminomethyl)cyclopropyl)acetonitrile instead of (S)-oxetane-2-ylmethaneamine and intermediate (0102-2). LCMS (ESI): m / z = 596.2 (M+H) + .

[0287] Example 28, (S)-2-((4-(6-((furan-2-ylmethyl)thio)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-28).

[0288]

[0289] The synthesis of compound 28 (0107-28) was performed according to Example 1, except that the left-hand fragment was replaced with furfuryl mercaptan instead of (4-chloro-2-fluorophenyl)methanol, and the remaining steps were the same. LCMS (ESI): m / z = 518.2 (M+H) + .

[0290] Example 29, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)-3,5-difluoropyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-29).

[0291]

[0292]

[0293] The synthesis of compound 29 (0107-29) is based on synthetic scheme seven, and the specific synthetic route is shown below.

[0294] 1. tBuOK,THF,rt.2,Pd2(dba)3,Xantphos,Cs2CO 3, 1,4-dioxane, 120℃ 3,3-bromopropene, Cs₂CO₃, DMF, 70℃ 4, Potassium osmium dihydrate, NaIO₄, rt 5, Stage 1: NH₂NHBOC, DCE, rt, Stage 2: NaBCNH₃ / NaB(CH₃COO)₃H, rt 6, 4M HCl in 1,4-dioxane, rt 7, Methyl orthoformate, AcOH 8, 4M HCl (aq.), DCM / CH₃OH = 1:1, 9, K₂CO₃, KI, Acetonitrile, 80℃, 9, NaOH, THF / CH₃OH / H₂O = 3:3:1

[0295] The synthesis steps of intermediate (0707-1) are as follows:

[0296] Step 1: 2 g (13.24 mmol) of 3-fluoro-4-(hydroxymethyl)benzonitrile (0301-2) was dissolved in 30 ml of anhydrous THF. 1.93 g (17.2 mmol) of potassium tert-butoxide was added under ice bath conditions. Then, 2.79 g (13.24 mmol) of 2-bromo-3,5,6-trifluoropyridine dissolved in 20 ml of anhydrous THF was added dropwise at room temperature. The reaction was carried out for about 3-5 hours under TLC monitoring. After the reaction was completed, the reaction was quenched with saturated brine. The sample was extracted three times with ethyl acetate and washed once with saturated brine. The filtrate was dried with magnesium sulfate and filtered. The sample was then mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 3.58 g (10.46 mmol) of 4-((6-bromo-3,5-difluoropyridin-2-yl)oxy)methyl)-3-fluorobenzonitrile (0701-1). LCMS(ESI): m / z = 342.0(M+H) + .

[0297] Step 2: 3.58 g (10.46 mmol) of 4-((6-bromo-3,5-difluoropyridin-2-yl)oxy)methyl)-3-fluorobenzonitrile (0701-1), (20.9 mmol) of tert-butyl carbamate were added to a sealing tube and dissolved in 70 ml of 1,4-dioxane solution. 1.44 g (1.57 mmol) of catalyst Pd2(dba)3, 1.82 g (3.14 mmol) of ligand Xantphos, and 8.5 g (26.14 mmol) of Cs2CO3 were added. The mixture was purged with N2 for 5 minutes, then reacted overnight in a sealed tube at 120 °C. After cooling to room temperature, the insoluble residue was filtered off. The filter cake was washed twice with ethyl acetate. The filtrate was dried over magnesium sulfate and filtered, then rinsed with 100-200 ml of ethyl acetate solution. The sample was mixed with silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 2.78 g (7.32 mmol) of tert-butyl(6-((4-cyano-2-fluorobenzyl)oxy)-3,5-difluoropyridin-2-yl)carbamate (0702-1). LCMS (ESI): m / z = 379.11 (M+H) + .

[0298] Step 3: 2.78 g (7.32 mmol) tert-butyl(6-((4-cyano-2-fluorobenzyl)oxy)-3,5-difluoropyridin-2-yl) carbamate was dissolved in 30 ml of N,N-dimethylformamide, and 3.04 g (21.97 mmol) of anhydrous potassium carbonate and 1.9 ml (21.97 mmol) of 3-bromopropene were added. The reaction was carried out in an oil bath at 70 °C for about 3 hours, and the reaction was monitored by LCMS. After the reaction was completed, the filtrate was extracted three times with ethyl acetate, washed three times with sodium chloride saturated brine, dried with magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain about 2.91 g tert-butylallyl(6-((4-cyano-2-fluorobenzyl)oxy)-3,5-difluoropyridin-2-yl) carbamate (0703-1).

[0299] LCMS(ESI): m / z = 419.15(M+H) + .

[0300] Step 4: Dissolve 2.91 g (6.95 mmol) tert-butylallyl (6-((4-cyano-2-fluorobenzyl)oxy)-3,5-difluoropyridin-2-yl)carbamate (0703-1) in 50 ml of a 1:1 mixture of tetrahydrofuran and water, and add potassium osmium tetroxide (VI). 25.6 mg (0.0695 mmol) of sodium periodate dihydrate and 7.44 g (34.75 mmol) of sodium periodate were reacted at room temperature for about 1 hour, resulting in the precipitation of a large amount of white solid. TLC monitoring was performed. After the reaction was complete, approximately 100 mL of saturated sodium thiosulfate aqueous solution was added, and the mixture was stirred for about 10 minutes. The mixture was extracted three times with ethyl acetate, and the organic layer was dried over magnesium sulfate. The mixture was filtered, and the filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 1.51 g of tert-butyl(6-((4-cyano-2-fluorobenzyl)oxy)-3,5-difluoropyridin-2-yl)(2-oxoethyl)carbamate (0704-1). LCMS (ESI): m / z = 421.12 (M+H) + .

[0301] Step 5, 1.51 g (3.59 mmol) tert-butyl(6-((4-cyano-2-fluorobenzyl)oxy)-3,5-difluoropyridin-2-yl)(2- Oxyethyl)carbamate (0704-1) was dissolved in 30 ml of 1,2-dichloroethane, and 0.95 g (7.2 mmol) of tert-butoxycarbamate was added. The reaction was carried out at room temperature for about 1 hour, monitored by TLC. After the reaction was completed, 2.28 g (10.77 mmol) of sodium triacetoxyborohydride and 677 mg (10.77 mmol) of sodium cyanoborohydride were added. The reaction was carried out for about 3 hours, monitored by LCMS. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate aqueous solution, extracted twice with dichloromethane, dried with magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain about 1.392 g of tert-butyl 2-(2-((tert-butoxycarbonyl)(6-((4-cyano-2-fluorobenzyl)oxy)-3,5-difluoropyridin-2-yl)amino)ethyl)hydrazine-1-carboxylic acid ester (0705-1). LCMS(ESI): m / z = 537.2(M+H) + .

[0302] Step 6: 1.392 g (2.592 mmol) of tert-butyl-2-(2-((6-bromopyridin-2-yl)(tert-butoxycarbonyl)amino)ethyl)hydrazine-1-carboxylic acid ester (0705-1) was dissolved in 24 mL of dichloromethane. 12 mL of a 4M solution of 1,4-dioxane with hydrogen chloride was added. The reaction was allowed to proceed for approximately 5 hours, resulting in the precipitation of a large amount of white solid. The reaction was monitored by LCMS. After the reaction was complete, the solvent was directly evaporated to obtain the hydrochloride salt of 4-((3,5-difluoro-6-((2-hydrazineethyl)amino)pyridin-2-yl)oxy)methyl)-3-fluorobenzonitrile (0706-1), which can proceed directly to the next step without purification. LCMS (ESI): m / z = 337.12 (M+H) + .

[0303] Step 7: Dissolve the hydrochloride salt (0706-1) of 4-((3,5-difluoro-6-((2-hydrazinoethyl)amino)pyridin-2-yl)oxy)methyl)-3-fluorobenzonitrile (0706-1) from the previous step in 24 ml of glacial acetic acid, add 12 ml of trimethyl orthoformate, and react overnight at 100°C under nitrogen protection. Evaporate the solvent using a rotary evaporator, add saturated sodium bicarbonate aqueous solution, extract three times with ethyl acetate, dry the organic layer with magnesium sulfate, filter, and mix the filtrate with 100-200 mesh silica gel powder. Analyze the filtrate using Flash-LC column chromatography to obtain approximately 784 mg (2.09 mmol) of 4-((3,5-difluoro-6-(1-formyl-5,6-dihydro-1,2,4-triazine-4(1H)-yl)pyridin-2-yl)oxy)methyl)-3-fluorobenzonitrile (0208-29). LCMS(ESI): m / z = 375.1(M+H) + .

[0304] Subsequent steps, referring to steps 7 and 9 of Example 1, finally yielded compound 29 (0107-29) (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)-3,5-difluoropyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-29). LCMS (ESI): m / z = 591.2 (M+H) + .

[0305] Example 30, (S)-2-((4-(2-((4-cyano-2-fluorobenzyl)oxy)pyrimidin-4-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-30).

[0306]

[0307] The synthesis of compound 30 (0107-30) was performed with reference to Example 29, replacing intermediate (0701-1) in Example 29 with intermediate 4-((4-bromopyrimidin-2-yl)oxy)methyl)-3-fluorobenzonitrile (0701-2). The synthesis of intermediate (0701-2) was performed with reference to the synthesis of compound Intermediate I-76 in patent US20210171499. LCMS (ESI): m / z = 556.2 (M+H) + .

[0308] Example 31, (S)-2-((4-(4-((4-cyano-2-fluorobenzyl)oxy)pyrimidin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-31).

[0309]

[0310] The synthesis of compound 31 (0107-31) was performed with reference to Example 29, except that intermediate 4-((2-chloropyrimidin-4-yl)oxy)methyl)-3-fluorobenzonitrile (0701-3) was used instead of intermediate (0701-1) in Example 29. The synthesis of intermediate (0701-3) was performed with reference to the synthesis of compound Intermediate I-4 in patent US20210171499. LCMS (ESI): m / z = 556.2 (M+H) + .

[0311] Example 32, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyrazin-2-yl)-5,6-dihydro-1,2,4-triazin-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-32).

[0312]

[0313] The synthesis of compound 32 (0107-32) was performed according to Examples 1 and 5, except that the starting material (0201-1) in Example 1 was replaced with (6-chloropyrazin-2-yl)carbamate tert-butyl ester. The remaining steps were similar, ultimately yielding (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyrazin-2-yl)-5,6-dihydro-1,2,4-triazin-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-32). LCMS (ESI): m / z = 556.2 (M+H) + .

[0314] Example 33, (S)-2-((4-(6-((5-chloropyridin-2-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid (0107-33).

[0315]

[0316] The synthesis of compound 33 (0107-33) was performed according to Example 1, in which (5-chloropyridin-2-yl)methanol (0301-10) was used instead of (4-chloro-2-fluorophenyl)methanol (0301-1). LCMS (ESI): m / z = 547.2 (M+H) + .

[0317] Example 34, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-thiophene[2,3-d]imidazol-5-carboxylic acid (0107-34).

[0318]

[0319] The synthesis of compound 34 (0107-34) was performed according to Example 5, wherein the synthesis of the right-hand intermediate (S)-2-(chloromethyl)-1-(oxetane-2-ylmethyl)-1H-thieno[2,3-d]imidazolium-5-carboxylic acid methyl ester (0105-9) was performed according to patent WO2021249492, replacing intermediate (0105-1). The remaining steps were the same, ultimately yielding (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-thieno[2,3-d]imidazolium-5-carboxylic acid (0107-34). LCMS (ESI): m / z = 561.2 (M+H) + .

[0320] Example 35, (S)-2-((4-(6-((4-chloro-3-methylbenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-35).

[0321]

[0322] The synthesis of compound 35 (0107-35) is described in Example 1. (4-chloro-3-methylbenzofuran-7-yl)methanol (0301-12) is used instead of (4-chloro-2-fluorophenyl)methanol (0301-1), and the synthetic route for (4-chloro-3-methylbenzofuran-7-yl)methanol (0301-12) is as follows:

[0323]

[0324] Step 1: Under nitrogen protection, 2.0 g (10.75 mmol) of methyl 4-chloro-2-hydroxybenzoate, 1.754 g (12.9 mmol) of bromoacetone, and 2.23 g (16.13 mmol) of potassium carbonate were added to N,N-dimethylformamide. The mixture was stirred at room temperature for 1 hour under TLC monitoring. After the reaction was complete, water was added, and a solid precipitated. The mixture was filtered, the residue was washed with water, and dried to obtain 2.1 g (8.68 mmol) of crude methyl 4-chloro-2-(2-oxopropoxy)benzoate. This product did not require further purification and was used directly in the next step.

[0325] Step 2: Under nitrogen protection, a mixture of 2.1 g (8.68 mmol) of methyl 4-chloro-2-(2-oxopropoxy)benzoate and 4.38 g (13 mmol) of polyphosphoric acid in dichloroethane was refluxed overnight. After cooling to room temperature, the reaction was quenched with water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel and subjected to Flash-LC column chromatography to obtain approximately 818 mg (3.65 mmol) of methyl 4-chloro-3-methylbenzofuran-7-carboxylic acid. LCMS (ESI): m / z = 224.02 (M+H) + .

[0326] Step 3: 818 mg (3.65 mmol) of methyl 4-chloro-3-methylbenzofuran-7-carboxylic acid was dissolved in 20 ml of anhydrous THF. Under nitrogen protection, the mixture was placed in an ice bath at 0 °C. 2.3 ml (5.52 mmol) of 2.4 M LiAlH4 THF solution was added dropwise. The reaction was carried out for half an hour and monitored by TLC. The reaction solution was added dropwise to ice water. After adding 30 ml of ethyl acetate, the insoluble matter was filtered through diatomaceous earth. The filter cake was washed with 20 ml of ethyl acetate and then extracted multiple times with ethyl acetate. The filtrate was dried over anhydrous magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 690 mg (3.52 mmol) of (4-chloro-3-methylbenzofuran-7-yl)methanol (0301-12).

[0327] Subsequent reactions, as described in Example 1, yielded compound 35 (0107-35). LCMS (ESI): m / z = 600.2 (M+H) + .

[0328] Example 36, 2-((4-(2-(5-chloropyridin-2-yl)-2-methylbenzo[d][1,3]dioxo-4-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (0107-36).

[0329]

[0330] The synthesis of compound 36 (0107-36) is based on synthetic scheme four, method one in scheme five, and scheme six. The specific synthetic routes are as follows.

[0331]

[0332]

[0333] The synthetic steps of the intermediate compound (S)-2-((4-(2,3-dihydroxyphenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid methyl ester (0411-1) are as follows:

[0334] Step 1: 5 g (26.6 mmol) of 3-bromocatechol was dissolved in 120 ml of DMF, 11.03 g (79.8 mmol) of K2CO3 was added, followed by 9.95 g (58.52 mmol) of benzyl bromide. The reaction was carried out at 60 °C and monitored by TLC. After the reaction was complete, the mixture was poured into water and extracted three times with ethyl acetate. The mixture was then washed three and four times with saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate. The filtrate was mixed with 100-200 mesh silica gel and subjected to flash-LC column chromatography to obtain approximately 9 g (24.47 mmol) of ((3-bromo-1,2-phenyl)bis(oxy)bis(methylene)diphenyl (0402-1).

[0335] Step 2: 9 g (24.47 mmol) of ((3-bromo-1,2-phenyl)bis(oxy)bis(methylene)diphenyl (0402-1) was added to a sealing tube, dissolved in 140 ml of 1,4-dioxane solution, and 3.37 g (3.67 mmol) of catalyst Pd2(dba)3 was added. 4.255 g (7.341 mmol) of the ligand Xantphos and 19.93 g (61.18 mmol) of Cs₂CO₃ were reacted under nitrogen for 5 minutes, then in a sealed tube at 120 °C overnight. After cooling to room temperature, the insoluble residue was filtered off, the filter cake was washed twice with ethyl acetate, the filtrate was dried over magnesium sulfate, filtered, and then mixed with 100-200 mesh silica gel powder. Flash-LC column chromatography yielded approximately 6.93 g (17.1 mmol) of tert-butyl(2,3-bis(benzyloxy)phenyl)carbamate (0403-1). LCMS (ESI): m / z = 405.2 (M+H) + .

[0336] Step 3: 6.93 g (17.1 mmol) of tert-butyl(2,3-bis(benzyloxy)phenyl)carbamate (0403-1) was dissolved in 50 mL of N,N-dimethylformamide, and 7.09 g (51.3 mmol) of anhydrous potassium carbonate and 4.43 mL (51.3 mmol) of 3-bromopropene were added. The mixture was reacted in an oil bath at 70 °C for approximately 3 hours, monitored by LC-LC. After the reaction, the filtrate was extracted three times with ethyl acetate, washed three times with saturated sodium chloride brine, dried over magnesium sulfate, and filtered. The filtrate was mixed with 100-200 mesh silica gel and subjected to Flash-LC column chromatography to obtain approximately 6.99 g of tert-butylallyl(2,3-bis(benzyloxy)phenyl)carbamate (0404-1). LC-LC (ESI): m / z = 445.2 (M+H) + .

[0337] Step 4: 6.99 g (15.7 mmol) tert-butylallyl (2,3-bis(benzyloxy)phenyl)carbamate (0404-1) was dissolved in 100 ml of a 1:1 mixture of tetrahydrofuran and water. 57.8 mg (0.157 mmol) potassium (VI) osmium tetroxide dihydrate and 16.8 g (78.5 mmol) sodium periodate were added. The reaction was carried out at room temperature for about 1 hour, during which a large amount of white solid precipitated. The reaction was monitored by TLC. After the reaction was completed, about 200 ml of saturated sodium thiosulfate aqueous solution was added. After stirring for about 10 minutes, the mixture was extracted three times with ethyl acetate. The organic layer was dried with magnesium sulfate and filtered. The filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain about 3.58 g tert-butyl (2,3-bis(benzyloxy)phenyl)(2-oxyethyl)carbamate (0405-1). LCMS (ESI): m / z = 447.2(M+H) + .

[0338] Step 5: 3.58 g (8.01 mmol) tert-butyl(2,3-bis(benzyloxy)phenyl)(2-oxyethyl)carbamate (0405-1) was dissolved in 70 ml of 1,2-dichloroethane. 2.11 g (16 mmol) of tert-butoxyformylhydrazine was added, and the reaction was carried out at room temperature for about 1 hour under TLC monitoring. After the reaction was completed, 5.08 g (24 mmol) of sodium triacetoxyborohydride and 1.508 g (24 mmol) of sodium cyanoborohydride were added, and the reaction was carried out for about 3 hours under LCMS monitoring. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate aqueous solution, extracted twice with dichloromethane, dried the organic layer with magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 3.2 g of 2-(2-((2,3-bis(benzyloxy)phenyl)(tert-butoxycarbonyl)amino)ethyl)hydrazine-1-carboxylic acid tert-butyl ester (0406-1). LCMS(ESI): m / z = 563.3(M+H)+ .

[0339] Step 6: 3.2 g (5.68 mmol) of 2-(2-((2,3-bis(benzyloxy)phenyl)(tert-butyloxycarbonyl)amino)ethyl)hydrazine-1-carboxylic acid tert-butyl ester (0406-1) was dissolved in 48 mL of dichloromethane. 24 mL of a 4M solution of 1,4-dioxane with hydrogen chloride was added. The reaction was allowed to proceed for approximately 5 hours, during which a large amount of white solid precipitated. The reaction was monitored by LCMS. After the reaction was complete, the solvent was directly evaporated to obtain the hydrochloride salt of 2,3-bis(benzyloxy)-N-(2-hydrazylethyl)aniline (0407-1), which can proceed directly to the next step without purification. LCMS (ESI): m / z = 363.2 (M+H) + .

[0340] Step 7: Dissolve the hydrochloride salt of 2,3-bis(benzyloxy)-N-(2-hydrazinoethyl)aniline (0407-1) from the previous step in 48 ml of glacial acetic acid, add 24 ml of trimethyl orthoformate, and react overnight at 100°C under nitrogen protection. Evaporate the solvent using a rotary evaporator, add saturated sodium bicarbonate aqueous solution, and extract three times with ethyl acetate. Dry the organic layer with magnesium sulfate, filter, and mix the filtrate with 100-200 mesh silica gel. Perform Flash-LC column chromatography to obtain approximately 1.64 g (4.09 mmol) of 4-(2,3-bis(benzyloxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-carbonaldehyde (0408-1). LCMS (ESI): m / z = 401.2 (M+H) + .

[0341] Step 8: 1.64 g (4.09 mmol) of 4-(2,3-bis(benzyloxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-carbonaldehyde (0408-1) was dissolved in 60 ml of a 1:1 (v / v) mixture of dichloromethane and methanol. Under ice bath conditions, 4 ml (approximately 16 mmol) of 4M hydrochloric acid aqueous solution was added, and the mixture was reacted at room temperature overnight. After overnight reaction, a large amount of saturated sodium bicarbonate aqueous solution was added, and the mixture was extracted with dichloromethane. The organic layer was dried over magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 1.38 g (3.7 mmol) of 4-(2,3-bis(benzyloxy)phenyl)-1,4,5,6-tetrahydro-1,2,4-triazine (0409-1). LCMS(ESI): m / z = 373.2(M+H) + .

[0342] Step 9: Add 1.38 g (3.7 mmol) of 4-(2,3-bis(benzyloxy)phenyl)-1,4,5,6-tetrahydro-1,2,4-triazine (0409-1), 1.534 g (11.1 mmol) of potassium carbonate, 112 mg (0.74 mmol) of potassium iodide, and 1.31 g (4.44 mmol) of (S)-2-(chloromethyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester. Add 70 ml of acetonitrile and react at 80 °C for approximately 3 hours, monitored by LC-MS. After the reaction is complete, filter the solution. Mix the filtrate with 100-200 mesh silica gel powder and precipitate by Flash-LC column chromatography to obtain approximately 1.52 g of (S)-2-((4-(2,3-bis(benzyloxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid methyl ester (0410-1). LC-MS (ESI): m / z = 631.3 (M+H) + .

[0343] Step 10: 1.52 g of (S)-2-((4-(2,3-bis(benzyloxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H benzo[d]imidazolium-6-carboxylic acid methyl ester (0410-1) was dissolved in 70 ml of methanol, and 150 mg of Pd / C (10% wt% Pd) was added. The reaction was carried out in H2 environment at room temperature and monitored by TLC. After the reaction was completed, the catalyst was filtered off, and the organic solvent was evaporated to dryness using a rotary evaporator to obtain approximately 1.08 g of (S)-2-((4-(2,3-dihydroxyphenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H benzo[d]imidazolium-6-carboxylic acid methyl ester (0411-1). LCMS(ESI): m / z = 451.2(M+H) + .

[0344] The intermediate (0411-1) then reacts with 5-chloro-2-ynylpyridine.

[0345]

[0346] Step 1: 1.08 g (2.39 mmol) of (S)-2-((4-(2,3-dihydroxyphenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1Hbenzimidazole-6-carboxylic acid methyl ester (0411-1) and 328 mg (2.39 mmol) of 5-chloro-2-ynylpyridine were added to a sealed tube, followed by 70 ml of toluene. The tube was then purged with N2 for 3 minutes, and finally 46 mg (0.0717 mmol) of the catalyst dodecacarbonyltriruthenium (Ru3(CO)) was added. 12 The mixture was further purged with N2 for 2 minutes and reacted overnight at 100°C. After cooling overnight, the insoluble catalyst was filtered off, and the sample was mixed with 100-200 mesh silica gel. Flash-LC column chromatography yielded 430 mg of methyl 2-((4-(2-(5-chloropyridin-2-yl)-2-methylbenzo[d][1,3]dioxo-4-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (0412-1). LCMS (ESI): m / z = 588.2 (M+H) + .

[0347] Step 2, 430 mg (0.731 mmol) of 2-((4-(2-(5-chloropyridin-2-yl)-2-methylbenzo[d][1,3]dioxo-4-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d] Imidazole-6-carboxylic acid methyl ester (0412-1) was dissolved in 35 ml of a 3:3:1 mixture of tetrahydrofuran / methanol / water. 88 mg (2.19 mmol) of NaOH was added, and the reaction was carried out at room temperature for 3–5 hours under TLC monitoring. After the reaction was complete, 30 g of water was added, and the organic solvent was evaporated to dryness by rotary evaporation. The pH was carefully adjusted to 6–7 with 1 M hydrochloric acid aqueous solution, and a large amount of NaCl was added to make the solution supersaturated. The solution was then extracted multiple times with a 4:1 mixture of ethyl acetate / methanol. The organic layer was dried over anhydrous magnesium sulfate, filtered, and then purified with 100–200 ml of water. The sample was mixed with silica gel and subjected to Flash-LC column chromatography to obtain 2-((4-(2-(5-chloropyridin-2-yl)-2-methylbenzo[d][1,3]dioxo-4-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-36). LCMS (ESI): m / z = 574.2 (M+H) + .

[0348] Example 37, 2-((4-(2-(4-chloro-2-fluorophenyl)-2-toluidine[d][1,3]dioxo-4-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-37).

[0349]

[0350] The synthesis of compound 37 (0107-37) is based on Example 36, but the intermediate (0411-1) was synthesized using methods two and six of synthesis scheme five to obtain the final product, as detailed below.

[0351]

[0352] Step 1: 1 g (2.22 mmol) of intermediate (0411-1) and 402 mg (2.33 mmol) of 4'-chloro-2'-fluoroacetophenone were added to 60 ml of toluene, followed by 43 mg (0.222 mmol) of p-toluenesulfonic acid monohydrate. The mixture was reacted overnight at 160 °C using a water separator. After cooling to room temperature, the mixture was mixed with 100-200 mesh silica gel and subjected to flash-LC column chromatography to obtain approximately 239 mg of methyl 2-((4-(2-(4-chloro-2-fluorophenyl)-2-methylbenzo[d][1,3]dioxo-4-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0412-2).

[0353] LCMS(ESI): m / z = 605.2(M+H) + .

[0354] Step 2: 239 mg (0.395 mmol) of 2-((4-(2-(4-chloro-2-fluorophenyl)-2-methylbenzo[d][1,3]dioxo-4-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate methyl ester (0412-2) was dissolved in 35 ml of a methanol / tetrahydrofuran / water mixture of 3:3:1. 48 mg of [the solution / concentrate] was then added. (1.2 mmol) NaOH was added, and the reaction was monitored by TLC. After completion, 30 mL of water was added, the organic solvent was evaporated to dryness, and the pH was carefully adjusted to 6-7 with 1 M hydrochloric acid aqueous solution. Extraction was performed with ethyl acetate / methanol = 3:1 and saturated sodium chloride. The organic phase was dried over anhydrous magnesium sulfate. The filtrate was mixed with 100-200 mesh silica gel and subjected to flash-LC column chromatography to obtain 122 mg of 2-((4-(2-(4-chloro-2-fluorophenyl)-2-methylbenzo[d][1,3]dioxo-4-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-37). LCMS (ESI): m / z = 591.2 (M+H) + .

[0355] Example 38, (S)-2-((4-(6-((4-chloro-2,3-dihydrobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-3-(oxetane-2-ylmethyl)-3H-imidazolium[4,5-b]pyridine-5-carboxylic acid (0107-38).

[0356]

[0357] The synthesis of compound 38 (0107-38) was performed according to Example 18, wherein the right-hand intermediate (S)-2-(chloromethyl)-3-(oxetane-2-ylmethyl)-3H-imidazolium[4,5-b]pyridine-5-carboxylic acid methyl ester was prepared according to the synthesis of intermediate compound 0116-1 of CN202110839013.8. LCMS (ESI): m / z = 589.2 (M+H) + .

[0358] Example 39, (S)-2-((4-(6-((4-cyano-2,3-dihydrobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-39).

[0359]

[0360] The synthesis of compound 39 (0107-39) is described in Example 1, wherein the synthesis of the left fragment 7-(hydroxymethyl)-2,3-dihydrobenzofuran-4-nitrile (0301-14) is as follows.

[0361]

[0362] The synthesis of 7-(hydroxymethyl)benzofuran-4-onitrile was performed according to intermediate (0201-1) in CN202110839013.8. Then, following the synthesis of intermediate (0301-8) in Example 18, 7-(hydroxymethyl)-2,3-dihydrobenzofuran-4-onitrile (0301-14) was prepared. The remaining steps were the same as in Example 1, ultimately yielding (S)-2-((4-(6-((4-cyano-2,3-dihydrobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-39). LCMS (ESI): m / z = 579.2 (M+H) + .

[0363] Example 40, (S)-2-((4-(6-((4-chloro-2,3-dihydrobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-4-fluoro-1H-benzo[d]imidazolium-6-carboxylic acid (0107-40).

[0364]

[0365] The synthesis of compound 40 (0107-40) was performed according to Examples 2 and 18, LCMS (ESI): m / z = 606.2 (M+H). + .

[0366] Example 41, (S)-2-((4-(6-((4-cyano-2,3-dihydrobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0107-41).

[0367]

[0368] The synthesis of compound 41 (0107-41) was performed according to Examples 2 and 39, LCMS (ESI): m / z = 597.2 (M+H). + .

[0369] Example 42, 2-((4-(6-((4-cyano-2-methyl-2,3-dihydrobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-((S)-oxetane-2-yl)methyl)-1Hbenzo[d]imidazolium-6-carboxylic acid (0107-42).

[0370]

[0371] The synthesis of compound 42 (0107-42) is described in Examples 2 and 39. LCMS (ESI): m / z = 611.2 (M+H) + .

[0372] Example 43, (S)-2-((4-(6-((4-cyano-2-(difluoromethoxy)benzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1Hbenzi[d]imidazolium-6-carboxylic acid (0107-43).

[0373]

[0374] The synthesis route is as described in Example 9. LCMS(ESI): m / z = 621.2(M+H) + .

[0375] Example 44, (S)-2-((4-(6-((4-chloro-2-methoxybenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-methyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0376]

[0377] The synthesis route is as described in Example 19. LCMS(ESI): m / z = 634.2(M+H) + .

[0378] Example 45, (S)-2-((4-(6-((4-chloro-2-fluorobenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-methyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0379]

[0380] The synthesis route is as described in Example 19. LCMS(ESI): m / z = 622.2(M+H) + .

[0381] Example 46, (S)-2-(4-(6-((4-chloro-2-(difluoromethoxy)benzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-methyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0382]

[0383] The synthesis route is as described in Example 9. LCMS(ESI): m / z = 630.2(M+H) + .

[0384] Example 47, (S)-2-(4-(6-((4-chloro-2-methylbenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0385]

[0386] The synthesis of compound 47 (0107-17) is described in Example 1. (4-chloro-2-methylbenzofuran-7-yl)methanol was used instead of (4-chloro-2-fluorophenyl)methanol. The synthetic route for (4-chloro-2-methylbenzofuran-7-yl)methanol is as follows:

[0387]

[0388] Step 1: Under nitrogen protection, 2.0 g (10.75 mmol) of methyl 4-chloro-2-hydroxybenzoate, 1.52 g (12.9 mmol) of propargyl bromide, and 2.23 g (16.13 mmol) of potassium carbonate were added to N,N-dimethylformamide. The mixture was stirred at 60 °C for 2 hours and monitored by TLC. After the reaction was complete, the mixture was cooled to room temperature, quenched with water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 2.3 g (10.27 mmol) of methyl 4-chloro-2-(propargyl-1-oxy)benzoate. LCMS (ESI): m / z = 224.1 (M+H) + .

[0389] Step 2: 2.3 g (10.27 mmol) of methyl 4-chloro-2-(prop-2-yn-1-oxy)benzoate was dissolved in N,N-diethylaniline (10 mL), and 1.71 g of cesium fluoride (11.26 mmol) was added. The reaction was carried out at 220 °C for 3 hours under nitrogen protection. After cooling to room temperature, the reaction solution was diluted with ethyl acetate, washed three times with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel and subjected to Flash-LC column chromatography to obtain approximately 1.05 g (4.69 mmol) of methyl 4-chloro-2-methylbenzofuran-7-carboxylic acid. LCMS (ESI): m / z = 224.1 (M+H) + .

[0390] Step 3: 1.05 g (4.69 mmol) of methyl 4-chloro-2-methylbenzofuran-7-carboxylic acid was dissolved in 20 mL of anhydrous THF. Under nitrogen protection, the mixture was placed in an ice bath at 0 °C. 1.95 mL (4.69 mmol) of 2.4 M LiAlH4 THF solution was added dropwise. The reaction was carried out for half an hour and monitored by TLC. The reaction solution was added dropwise to ice water. After adding 30 mL of ethyl acetate, the insoluble matter was filtered through diatomaceous earth. The filter cake was washed with 20 mL of ethyl acetate and then extracted multiple times with ethyl acetate. The filtrate was dried over anhydrous magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 790 mg (4.02 mmol) of (4-chloro-2-methylbenzofuran-7-yl)methanol.

[0391] Subsequent reactions, as described in Example 1, yielded compound 47 (0107-47). LCMS (ESI): m / z = 600.2 (M+H) + .

[0392] Example 48, (S)-2-(4-(6-((4-cyano-2-ethoxybenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-1-(oxetane-2-methyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0393]

[0394] The synthesis route is as described in Example 9. LCMS(ESI): m / z = 581.3(M+H) + .

[0395] Example 49, (S)-2-(4-(6-((4-chloro-2-methylbenzofuran-7-yl)methoxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0396]

[0397] The synthesis route is as described in Example 47. LCMS(ESI): m / z = 618.2(M+H) + .

[0398] Example 50, (S)-2-(4-(6-(4-cyano-2-ethoxybenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-ylmethyl)-4-fluoro-1-(oxetane-2-methyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0399]

[0400] The synthesis route is as described in Example 10. LCMS(ESI): m / z = 599.3(M+H) + .

[0401] Example 51, (S)-2-(4-(6-(4-chloro-2-methoxybenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-ylmethyl)-4-fluoro-1-(oxetane-2-methyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0402]

[0403] The synthesis route is as described in Example 10. LCMS(ESI): m / z = 594.2(M+H) + .

[0404] Example 52, (S)-2-(4-(3-((4-cyano-2-methoxybenzyl)oxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-ylmethyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0405]

[0406] The synthesis route is as follows:

[0407]

[0408] 1: (Boc)₂O, DMAP, THF; 2: 3-bromopropene, Cs₂CO₃, DMF; 3: NaIO₄, K₂O₅O₄·9H₂O, THF, H₂O; 4: tert-butoxyformylhydrazine, NaBH₃CN, sodium triacetoxyborohydride, DCE; 5: 4M hydrogen chloride (1,4-dioxane solution), DCM; 6: trimethyl orthoformate, glacial acetic acid; 7: H₂, Pd / C, MeOH.

[0409] Step 1: 6 g (30.14 mmol) of 3-(benzyloxy)aniline was dissolved in 100 mL of tetrahydrofuran, and 7.4 g (60.28 mmol) of DMAP was added. The mixture was heated to 70 °C, and then 7.24 g (33.15 mmol) of (Boc)₂O was added dropwise. The reaction was carried out at this temperature for 1 h. The reaction solution was mixed with 100-200 mesh silica gel powder, and Flash-LC column chromatography was used to obtain approximately 6 g (20.07 mmol) of tert-butyl (3-(benzyloxy)phenyl)carbamate. LCMS (ESI): m / z = 299.2 (M+H) + .

[0410] Step 2: 6 g (20.07 mmol) of tert-butyl (3-(benzyloxy)phenyl)carbamate was dissolved in 50 mL of N,N-dimethylformamide, and 13.09 g (40.14 mmol) of cesium carbonate and 4.86 g (40.14 mmol) of 3-bromopropene were added. The mixture was reacted overnight in an oil bath at 70 °C, monitored by LC-LC. After the reaction, the filtrate was extracted three times with ethyl acetate, washed three times with saturated sodium chloride brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 6.65 g (19.60 mmol) of allyl (3-(benzyloxy)phenyl)carbamate. LC-LC (ESI): m / z = 339.2 (M+H) + .

[0411] Step 3: 6.65 g (19.60 mmol) of allyl (3-(benzyloxy)phenyl)carbamate tert-butyl ester was dissolved in 150 mL of a 1:1 mixture of tetrahydrofuran and water. 305 mg (0.98 mmol) of potassium (VI) osmium tetroxide dihydrate and 12.47 g (58.8 mmol) of sodium periodate were added. The reaction was carried out at room temperature for about 1 hour, resulting in the precipitation of a large amount of white solid. TLC monitoring was performed. After the reaction was complete, approximately 100 mL of saturated sodium thiosulfate aqueous solution was added. After stirring for about 10 minutes, the mixture was extracted three times with ethyl acetate. The organic layer was dried over magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel and subjected to Flash-LC column chromatography to obtain approximately 4 g (11.72 mmol) of (3-(benzyloxy)phenyl)(2-oxyethyl)carbamate tert-butyl ester. LCMS (ESI): m / z = 341.2 (M+H) + .

[0412] Step 4: 4 g (11.72 mmol) of (3-(benzyloxy)phenyl)(2-oxyethyl)carbamate tert-butyl ester was dissolved in 50 mL of 1,2-dichloroethane. 3.10 g (23.44 mmol) of tert-butoxyformylhydrazine was added, and the reaction was carried out at room temperature for about 1 hour, monitored by TLC. After the reaction was completed, 7.45 g (35.16 mmol) of triacetoxyborohydride and 2.22 g (35.16 mmol) of cyanoborohydride were added, and the reaction was carried out for about 3 hours, monitored by LCMS. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate aqueous solution, extracted twice with dichloromethane, dried with magnesium sulfate, filtered, and the filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 3.6 g (7.64 mmol) of (3-(benzyloxy)phenyl)(2-(((tert-butoxycarbonyl)amino)methyl)amino)ethyl)carbamate tert-butyl ester. LCMS(ESI): m / z = 471.3(M+H) + .

[0413] Step 5: 3.6 g (7.64 mmol) of (3-(benzyloxy)phenyl)(2-(((tert-butoxycarbonyl)amino)methyl)amino)ethyl)carbamate tert-butyl ester was dissolved in 30 mL of dichloromethane, and 20 mL of a 4M solution of 1,4-dioxane hydrogen chloride was added. The reaction was allowed to proceed for approximately 5 hours, during which a large amount of white solid precipitated. The reaction was monitored by LCMS. After the reaction was complete, the solvent was directly evaporated to obtain N. 1 -(aminomethyl)-N 2 3-(benzyloxy)phenyl)ethane-1,2-diamine hydrochloride can proceed directly to the next reaction without purification. LCMS (ESI): m / z = 271.2 (M+H) + .

[0414] Step 6, take the product N from the previous step 1 -(aminomethyl)-N 2 3-(benzyloxy)phenyl)ethane-1,2-diamine hydrochloride was dissolved in 30 mL of glacial acetic acid, and 15 mL of trimethyl orthoformate was added. The mixture was reacted overnight at 100 °C under nitrogen protection. The solvent was evaporated to dryness using a rotary evaporator, and a saturated sodium bicarbonate aqueous solution was added. The mixture was extracted three times with ethyl acetate, and the organic layer was dried over magnesium sulfate. The mixture was filtered, and the filtrate was mixed with 100-200 mesh silica gel and subjected to Flash-LC column chromatography to obtain approximately 1.7 g (5.76 mmol) of 4-(3-(benzyloxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-formaldehyde. LCMS (ESI): m / z = 295.1 (M+H) + .

[0415] Step 7: Dissolve 1.7 g (5.76 mmol) of 4-(3-(benzyloxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-carboxaldehyde in 20 mL of methanol, add 170 mg of 10% Pd / C, and react under a hydrogen atmosphere for 3 hours. After the reaction is complete, filter and evaporate to dryness to obtain 1.14 g (5.54 mmol) of 4-(3-hydroxyphenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-carboxaldehyde. LCMS (ESI): m / z = 205.1 (M+H) + .

[0416] Step 8: 90 mg (0.44 mmol) of 4-(3-hydroxyphenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-formaldehyde was dissolved in 4 mL of N,N-dimethylformamide. 287 g (0.88 mmol) of cesium carbonate and 155 mg (0.66 mmol) of 4-(bromomethyl)-3-methoxybenzonitrile were added. The mixture was reacted overnight in an oil bath at 70 °C. The reaction was monitored by LC-MS. After the reaction, the filtrate was extracted three times with ethyl acetate, washed three times with saturated sodium chloride brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was mixed with 100-200 mesh silica gel powder and subjected to Flash-LC column chromatography to obtain approximately 150 mg (0.43 mmol) of 4-((3-(1-formyl-5,6-dihydro-1,2,4-triazine-4(1H)-yl)phenoxy)methyl)-3-methoxybenzonitrile. LCMS(ESI): m / z = 350.1(M+H) + .

[0417] Subsequent reactions, as described in Example 2, yielded compound 52 (0107-52). LCMS (ESI): m / z = 584.2 (M+H) + .

[0418] Example 53, (S)-2-(4-(3-((4-chloro-2-methoxybenzyl)oxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-ylmethyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0419]

[0420] The synthesis route is as described in Example 52. LCMS(ESI): m / z = 593.2(M+H) + .

[0421] Example 54, (S)-2-((4-(3-)((4-cyano-2,3-dihydrobenzofuran-7-yl)methoxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0422]

[0423] The synthesis route is as described in Example 52. LCMS(ESI): m / z = 596.2(M+H) + .

[0424] Example 55, (S)-2-((4-(3-)((4-chloro-2,3-dihydrobenzofuran-7-yl)methoxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0425]

[0426] The synthesis route is as described in Example 52. LCMS(ESI): m / z = 605.2(M+H) + .

[0427] Example 56, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-methyl-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0428]

[0429] The synthesis route is as described in Example 5. LCMS(ESI): m / z = 569.2(M+H) + .

[0430] Example 57, (S)-2-((4-(6-((4-trifluoromethyl-2-methoxybenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1Hbenzi[d]imidazolium-6-carboxylic acid

[0431]

[0432] The synthetic route was the same as in Example 2, except that (4-trifluoromethyl-2-methoxyphenyl)methanol was used instead of (4-chloro-2-fluorophenyl)methanol as the starting material. LCMS (ESI): m / z = 629.2 (M+H) + .

[0433] Example 58, (S)-2-((4-(6-((4-cyano-2-methoxybenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1Hbenzi[d]imidazolium-6-carboxylic acid methyl ester

[0434]

[0435] It was obtained as a synthetic intermediate for compound 10. LCMS (ESI): m / z = 600.2 (M+H) + .

[0436] Example 59, (S)-2-((4-(6-((4-cyano-2-methoxybenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1Hbenzi[d]imidazolium-6-carboxylic acid ethyl ester

[0437]

[0438] One equivalent of compound 10, 1.1 equivalents of iodoethane, and 1.5 equivalents of potassium carbonate were reacted in DMF overnight at room temperature. The DMF was evaporated under reduced pressure, and the residue was separated by preparative liquid chromatography. LCMS (ESI): m / z = 614.2 (M+H) + .

[0439] Example 60, (S)-2-(4-(2-(4-cyano-2-methoxybenzyl)oxy)phenyl)-5,6-dihydro-1,2,4-triazine-1(4H)-ylmethyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0440]

[0441] The synthetic route was the same as in Example 52, except that the starting material was 2-(benzyloxy)aniline instead of 3-(benzyloxy)aniline. LCMS (ESI): m / z = 584.2 (M+H) + .

[0442] Example 61, (S)-2-((4-(6-((4-cyano-2-methoxybenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1Hbenzi[d]imidazolium-6-carboxylic acid sodium salt

[0443]

[0444] One equivalent of compound 10 and one equivalent of sodium methoxide were added to methanol and stirred at room temperature for 30 minutes. Methanol was then distilled off under reduced pressure to give compound 61. LCMS (ESI): m / z = 585.2 (M+H) + . 1 H NMR (500MHz, DMSO) δ8.02(s,1H),7.86 (s,1H),7.66(t,J=8.0Hz,1H),7.52(d,J=1.5Hz,1H),7.51–7.50(m,1H),7.49(d,J= 1.5Hz,1H),7.42(dd,J=7.8,1.5Hz,1H),6.52(d,J=8.1Hz,1H),6.46(d,J=8.0Hz, 1H),5.36(s,2H),5.09–5.02(m,1H),4.81(dd,J=15.3,7.1Hz,1H),4.68(dd,J=15.4, 2.9Hz,1H),4.51–4.43(m,2H),4.40–4.30(m,2H),3.89(s,3H),3.74(t,J=5.1Hz,2H), 3.16–3.03(m,2H),2.73–2.64(m,1H),2.44–2.33(m,1H).

[0445] Example 62, 1,3-Dihydroxy-2-(hydroxymethyl)propyl-2-amine(S)-2-((4-(6-((4-cyano-2-methoxybenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-fluoro-1-(oxetane-2-ylmethyl)-1Hbenzi[d]imidazolium-6-carboxylate

[0446]

[0447] One equivalent of compound 10 and one equivalent of tris(hydroxymethyl)aminomethane were dissolved in methanol and stirred at room temperature for 30 minutes. The methanol was then distilled off under reduced pressure to give compound 62. LCMS (ESI): m / z = 585.2 (M+H) + . 1 H NMR(500MHz,DMSO)δ8.01(d, J=1.1Hz,1H),7.86(s,1H),7.65(t,J=8.0Hz,1H),7.51(d,J=1.5Hz,1H),7.51–7.49 (m,1H),7.48(d,J=1.5Hz,1H),7.41(dd,J=7.8,1.5Hz,1H),6.51(d,J=8.0Hz,1H), 6.45(d,J=8.0Hz,1H),5.35(s,2H),5.09–5.00(m,1H),4.80(dd,J=15.4,7.1Hz,1H), 4.67(dd,J=15.4,2.9Hz,1H),4.50–4.42(m,2H),4.39–4.29(m,2H),3.88(s,3H),3.7 3(t,J=5.1Hz,2H),3.43(s,6H),3.15–3.03(m,2H),2.72–2.63(m,1H),2.43–2.32(m, 1H).

[0448] Example 63, (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-yl)methyl)-4-ethyl-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid

[0449]

[0450] The synthesis route is as described in Example 5. LCMS(ESI): m / z = 583.2(M+H) + .

[0451] The compounds of this invention 1 H-NMR.

[0452]

[0453]

[0454]

[0455]

[0456]

[0457]

[0458]

[0459]

[0460] Example 64: cAMP level determination in hGLP-1R / HEK293 cell line

[0461] The human GLP-1R gene (hGLP-1R) was amplified from the cDNA of the human colorectal adenocarcinoma cell line NCl-H716 (Chinese Academy of Sciences Cell Bank, #TCHu210) and cloned into the HA-pcDNA3.1 vector (Addgene, #128034), positioning the HA-tag at the N-terminus of hGLP-1R. The hGLP-1R / HA-pcDNA3.1 plasmid was transfected into HEK293 cells to construct a stable hGLP-1R / HEK293 cell line. The hGLP-1R / HEK293 cells were digested, centrifuged, and resuspended in culture medium (DMEM + 0.2% BSA), and the cell density was adjusted to 2 × 10⁶ cells / year. 5 Cells / mL, add IBMX (3-isobutyl-1-methylxanthine, final concentration 0.5mM) to the cell suspension, at 5μL (1×10⁻⁶ cells / mL). 3 Cells were added per well to a 384-well plate. The compound was dissolved in DMSO to prepare a 10 mM stock solution. For experiments, a 100 μM concentration was used as the starting point, with 10-fold serial dilutions for a total of 7 spots. The 8th spot was diluted with DMSO. 2 μL of different concentrations of the test compound were added to 98 μL of culture medium, vortexed, and then 5 μL was added to each 384-well plate (final concentration of test compound: 0-100 nM; final concentration of DMSO: 1%). The plate was incubated in a dark incubator (37°C, 5% CO2) for 30 min. Then, 10 μL of cAMP detection reagent (LANCE Ultra cAMP Detection Kit, PerkinElmer, #TRF0264) was added, mixed thoroughly, and incubated at room temperature in the dark for 1 h. After the reaction, the HTRF signal was read using an Envision 2104 multi-functional microplate reader. All samples were tested in triplicate. The receptor-donor excitation signal ratio for each single well was calculated using the formula Ratio = Signal 665 nM / Signal 620 nM. The HTRF signal ratio and the corresponding compound concentration were analyzed using GraphPad Prism 8 software. The EC50 of the compound was calculated by fitting an S-shaped dose-response curve. 50 value.

[0462] Table 1. Effects of the compounds of the present invention on cAMP levels in the hGLP-1R / HEK293 cell line.

[0463]

[0464]

[0465] As can be seen from the table above, the compounds in this invention can activate hGLP-1R, and some compounds have higher activity than the control compound PF-06882961. Furthermore, the 5,6-dihydro-1,2,4-triazine structure makes the ClogP value of the compounds significantly different from that of the control compound PF-06882961.

[0466] Comparative data on GLP-1 receptor agonistic activity:

[0467] The distinguishing structural feature of the compounds represented by general formula (I) of this invention lies in the fact that the connecting heterocycle in the middle is 5,6-dihydro-1,2,4-triazine, which is different from other heterocycles disclosed in existing patent applications, such as 6-oxopyridazine (compound A), 2-oxopyrimidine (compound B), and 2-oxy-1,3-oxazine (compound C) as shown in Table 2 below. Existing publications and the inventors' test results both indicate that the effect of heterocycle transformation on the GLP-1 receptor agonist activity of the compounds is difficult to predict. Compared with compounds A, B, and C, which have similar structures, the compounds of this invention exhibit higher GLP-1 receptor agonist activity.

[0468] Table 2. Effects of comparative compounds on cAMP levels in hGLP-1R / HEK293 cell line

[0469]

[0470]

[0471] Example 65: cAMP level determination in mGLP-1R-S33W transiently transfected HEK293 cells

[0472] A mouse GLP-1R plasmid, mGLP-1R / HA-pcDNA3.1, was constructed, with the HA-tag positioned at the N-terminus of mGLP-1R. The 33rd serine (S) residue in mGLP-1R was mutated to tryptophan (W). The mGLP-1R-S33W / HA-pcDNA3.1 plasmid was prepared at a rate of 1 μg plasmid / 2 × 10⁻⁶ ppm. 6HEK293 cells were transfected at a ratio of [number] cells / well, and cultured in an incubator (37°C, 5% CO2) for 24 hours before subsequent experiments. The transfected cells were digested, centrifuged, and resuspended in culture medium (DMEM + 0.2% BSA). Subsequent cAMP assays were performed under the same conditions as in Experiment 1. All samples were tested in triplicate. The receptor-donor excitation signal ratio for each well was calculated using the formula Ratio = Signal 665 nM / Signal 620 nM. The HTRF signal ratio and corresponding compound concentration were analyzed using GraphPad Prism 8 software. The EC50 of the compound was calculated by fitting an S-shaped dose-response curve. 50 value.

[0473] Table 3. Effects of the compounds of the present invention on cAMP levels in mGLP-1R-S33W transiently transfected HEK293 cells.

[0474] compound <![CDATA[Determination of EC 50 (pM)]]> compound <![CDATA[Determination of EC 50 (pM)]]> PF-06882961 450 1 320 2 370 3 4370 4 300 5 1120 6 370 7 5840 8 380 9 260 10 50 11 2430 12 700 13 2000 14 380 15 360 16 90 17 250 18 260

[0475] As can be seen from the table above, the compounds in this invention can activate mGLP-1R-S33W, and some compounds have higher activity than the control compound PF-06882961.

[0476] Example 66: Evaluation of the blocking effect of the compound on hERG potassium channels

[0477] 1. Compound preparation

[0478] 1.1 Dilute the compound stock solution with DMSO. Take 10 μL of the compound stock solution and add it to 20 μL of DMSO solution. Dilute 3 times to 6 DMSO concentrations.

[0479] 1.2 Take 4 μL of each of the six DMSO concentrations and add them to 396 μL of extracellular fluid. Dilute 100-fold to six intermediate concentrations. Then take 80 μL of each of the six intermediate concentrations and add them to 320 μL of extracellular fluid. Dilute 5-fold to the final concentration to be tested.

[0480] 1.3 The highest tested concentration was 40.00 μM, and the following concentrations were 40.00, 13.33, 4.44, 1.48, 0.49, and 0.16 μM, for a total of 6 concentrations.

[0481] 1.4 The DMSO content in the final test concentration does not exceed 0.2%. This concentration of DMSO has no effect on the hERG potassium channel.

[0482] 1.5 Compound preparation: The entire dilution process was completed using a Bravo instrument.

[0483] 2. Qpatch Fully Automated Patch Clamp Test Results

[0484] 2.1 Structure of the reference compound for hERG potassium channel blocking assay

[0485] Test results and comparison data:

[0486] Both publicly available information and the inventors' test results indicate that when the intermediate linking heterocycle of a GLP-1 receptor agonist molecule is a piperidine ring, the compound often exhibits significant hERG potassium channel blocking activity. Examples include PF-06882961 shown in Table 4 and compound 3 disclosed in patent application CN113480534A, which inhibits the hERG channel's IC50. 50 Values ​​range from 2 to 6 μM. In medicinal chemistry, hERG channel blockade is considered an indicator of cardiotoxicity risk and should be avoided as much as possible. The distinguishing structural feature of the compound represented by general formula (I) of this invention is that the connecting heterocyclic ring in the middle is 5,6-dihydro-1,2,4-triazine, rather than a piperidine ring. Unexpectedly, the introduction of the 5,6-dihydro-1,2,4-triazine ring significantly weakens the blocking effect of the compound on the hERG channel; therefore, the compound of this invention has a lower risk of cardiotoxicity.

[0487]

[0488] Table 4: Detection results of the blocking effect of compounds on hERG potassium channels

[0489] compound <![CDATA[IC 50 (μM)]]> Reference compound F-06882961 2.62 Reference compound 3 (CN113480534A) 5.7 6 114.52 9 >40 10 >40 15 >40 16 >40 18 34.45 38 >40 39 >40 40 87.40 41 >40 45 >40 52 >40

[0490] Example 67: Pharmacodynamic Experiment

[0491] Genetically engineered hGLP-1R mice, male, 8-10 weeks old, specific pathogen-free (SPF) grade, were purchased from Biocytogen Jiangsu Gene Biotechnology Co., Ltd. All experimental animals were housed in an SPF-grade environment at the National Compound Sample Bank animal facility, with a temperature of 24±2℃, relative humidity of 40-60%RH, air cleanliness level 7, and a 12h / 12h day / night cycle. They were continuously provided with cobalt-60 radioactively sterilized complete pelleted rat feed (maintenance feed for mice and rats from Shanghai Shilin Biotechnology Co., Ltd.), with unlimited access. They also had access to tap water (sterilized by autoclaving) provided continuously and freely. The cages were transparent polyetherimide cages (Suzhou Fengshi Laboratory Animal Equipment Co., Ltd., CP-8 type mouse cages), and the bedding was corn cobs (Dezhou Gumei Agricultural Technology Co., Ltd., used after autoclaving). Each cage contained 3-5 animals, and the cage tags were labeled with the IACUC approval number, experiment number, experiment start time, person in charge, experimenters, animal origin, group, and animal number. Purchased mice were allowed to acclimatize to this environment for at least 7 days before being used in the experiment. The animal use method in this experiment was approved by the IACUC Committee of the Shanghai Institute of Materia Medica.

[0492] 1. Oral glucose tolerance test in hGLP-1R mice

[0493] (1) Mouse model: The above hGLP-1R KI mice, eight weeks old, 4 males in each experimental group.

[0494] (2) Preparation of compounds: vehicle group, ultrapure water containing 1% DMSO; PF-06882961, dosage 1 mg / kg, dissolved in a small amount of DMSO and then added to pure water; test compound, dosage 1 mg / kg, dissolved in a small amount of DMSO and then added to pure water.

[0495] (3) Experimental procedure: Mice were starved overnight, and blood glucose in the tail of the mice was measured. After oral administration of the drug for 60 minutes, blood glucose was measured again as the zero point blood glucose. Then, 2g / kg of sugar was administered orally. Blood glucose was measured using a blood glucose meter at 15 minutes, 30 minutes, 60 minutes, 90 minutes and 120 minutes after the administration of sugar.

[0496] (4) Experimental Results

[0497] like Figure 1 and Figure 2 As shown, both compound 10 of the present invention and the positive control PF-06882961 significantly reduced blood glucose concentration when administered orally at 1 mg / kg, demonstrating potential for the treatment of diabetes.

[0498] 2. Feeding inhibition experiment in hGLP-1R mice

[0499] (1) Mouse model: The above hGLP-1R KI mice, eight weeks old, 4 males in each experimental group.

[0500] (2) Compound preparation: vehicle group, ultrapure water, containing 10% DMSO + 1% Tween-80; PF-06882961, dosage 30mg / kg, dissolved in a small amount of DMSO and then added to the solvent; test compound, dosage 30mg / kg, dissolved in a small amount of DMSO and then added to the solvent.

[0501] (3) Experimental procedure: Mice were starved overnight. Each mouse was weighed in a cage. The drug was administered by gavage and added to the cage. The weight of food in each cage was weighed at 0, 2, 4, 8 and 24 hours after administration, and the food intake of the mice was calculated.

[0502] (4) Experimental Results

[0503] like Figure 3 As shown, compound 10 of the present invention significantly inhibited food intake in experimental mice within 2 to 24 hours after oral administration. It is known that commercially available peptide GLP-1 receptor agonists have effects on suppressing appetite and promoting weight loss; therefore, the compound of the present invention also has the potential for use in controlling appetite and promoting weight loss.

[0504] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound of general formula (I) or a pharmaceutically acceptable salt thereof: In the formula, ring A is phenyl or pyridinyl; R 1 Selected from the group consisting of: 4-membered heterocyclic substituted C1-C4 alkyl, C3-cycloalkyl substituted C1-C4 alkyl, 5-membered heteroaryl substituted C1-C4 alkyl, and C1-C4 alkoxy substituted C1-C4 alkyl; wherein the above C3-cycloalkyl and 5-membered heteroaryl groups are optionally substituted by one or more substituents selected from the group consisting of: C1-C4 alkyl or cyano-substituted C1-C4 alkyl; R 2 For H; R 3 For H; Each R 4 Independently H or halogen; R 5 For H; Q is ; in, Z 1 Z 2 Z 3 Each independently for CR 11 Or N; Z 4 Let N be the number of people in the group. Y is O; Ring B is a phenyl or The ring C is a 5-membered heteroaryl group or a 5-membered partially unsaturated heterocyclic group; W 1 and W 2 Selected from CR 12 ; Each R 6 Independently selected from the group consisting of: H, halogen, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy, and cyano; R 9 It can be H, halogen, or cyano; Each R 10 Independently, it is H, halogen, or C1-C3 alkyl; Each R 11 Independently H or halogen; R 12 For H; a is 0 or 1; m is 0 or 1; n is 1, 2, or 3; p is 0 or 1.

2. The compound according to claim 1, characterized in that, Z 1 Z 2 and Z 3 Each independently selected from CR 11 Or N, Z 4 Let N be the number of people in the group. Y is O; Ring B is phenyl; Each R 11 It can be H or halogen independently.

3. The compound according to claim 1, characterized in that, The compound has the structure shown in formula (II) or formula (III): Where: R 1 R 2 R 3 R 4 R 5 Q, a, and m are as defined above.

4. The compound according to claim 1, characterized in that, Z 1 Z 2 and Z 3 Each independently selected from CR 11 Z 4 For N; each R 11 It can be H, F, Cl, or Br independently.

5. The compound according to claim 1, characterized in that, Z 1 Z 2 and Z 3 Each can be independently designated as CH or CF.

6. The compound according to claim 1, characterized in that, Q is selected from: 、 、 、 、 or , in, R 6 R 9 R 10 R 11 n and p are as defined in claim 1; g can be 0, 1, 2, or 3.

7. The compound according to claim 1, characterized in that, Selected from the following group: , or , Among them, R 6 R 9 R 10 n and p are as defined in claim 1.

8. The compound according to claim 1, characterized in that, The ring C is selected from furanyl.

9. The compound according to claim 1, characterized in that, for or .

10. The compound according to claim 1, characterized in that, for or .

11. The compound according to claim 1, characterized in that, The compound has the structure shown in formula (IV), formula (V) or formula (VI): Where: R 1 R 4 R 5 R 6 R 9 R 10 R 11 m, n and p are as defined in claim 1; g is 0, 1, 2 or 3; X is selected from CH or N.

12. The compound according to claim 1, characterized in that, R 1 Selected from C1 alkyl groups substituted with 4-membered heterocyclic groups, C1 alkyl groups substituted with 5-membered heteroaryl groups, and C1-C4 alkyl groups substituted with C3 cycloalkyl groups; wherein the C3 cycloalkyl group and the 5-membered heteroaryl group are optionally substituted by one or more substituents selected from the group consisting of H, C1-C4 alkyl groups or cyano-substituted C1-C4 alkyl groups.

13. The compound according to claim 1, characterized in that, R 1 Selected from , or .

14. The compound according to claim 1, characterized in that, R 4 Each is independently selected from H, F, Cl, or Br.

15. The compound according to claim 1, characterized in that, R 6 Each is independently selected from H, F, Cl, Br, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy, and cyano.

16. The compound according to claim 1, characterized in that, R 9 Selected from F, Cl, Br or cyano groups.

17. The compound according to claim 1, characterized in that, R 10 Each is independently selected from H, F, Cl, Br or C1-C3 alkyl groups.

18. The compound according to claim 1, characterized in that, The compound is: or 。 19. A pharmaceutical composition, characterized in that, It comprises: a compound as described in any one of claims 1-18 or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

20. Use of the compound of any one of claims 1-18 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 19 in the preparation of a medicament for treating type II diabetes or obesity.

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